1-galactose derivatives

ABSTRACT

Disclosed are novel 1-galactose derivatives which inhibit binding of toxins, such as heat-labile enterotoxin or cholera toxin, to their receptors either in vitro or in vivo. Additionally, disclosed are compounds which inhibit binding of enterovirulent organisms (e.g., bacteria, virus, fungi, and the like), such as Vibrio cholerae and enterotoxigenic strains of Escherichia coli, to their cell surface receptors.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. Ser. No. 08/751,510,filed Nov. 15, 1996, which application claims the benefit of U.S.Provisional Application No. 60/030,794, filed Nov. 14, 1996, whichapplications are incorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

Field of the Invention

This invention relates to novel 1-galactose derivatives which inhibitbinding of toxins, such as heat-labile enterotoxin (LT) or cholera toxin(CT), to their receptors either in vitro or in vivo. Additionally, thecompounds of this invention inhibit binding of enterovirulent organisms(e.g., bacteria, virus, fungi, and the like), such as Vibrio choleraeand enterotoxigenic strains of Escherichia coli, to their cell surfacereceptors.

REFERENCES

The following publications, and patents are cited in this application assuperscript numbers:

¹ Spangler, B. D., "Structure and Function of Cholera Toxin and RelatedEscherichia coli Heat-Labile Enterotoxin", Microbiological Reviews,56(4):622-647 (1992).

² Hol, W. G. J., et al., "Structure and Function of E. coli Heat-LabileEnterotoxin and Cholera Toxin B Pentamer", Bacterial Toxins andVirulence Factors in Disease, Ed. by J. Moss et al., Marcel Dekker, Inc.(1995).

³ Williams (ed.), Synthesis of Optically Active α-Amino Acids, PergamonPress (1989).

⁴ Evans et al., J. Amer. Chem. Soc., 112:4011-4030 (1990).

⁵ Pu et al., J. Amer. Chem. Soc., 56:1280-1283 (1991).

⁶ Williams et al., J. Amer. Chem. Soc., 113:9276-9286 (1991).

⁷ Kagen et al., Synlett, 1990, 643-650.

⁸ E. Hasegawa, K. Ishiyama, T. Horaguchi, T. Shimizu, J. Org. Chem.1991, 56, 1631-1635.

⁹ H. Paulsen, K. Eberstein, W. Koebernick, Tetrahedron Letters, 45-50,4377-4380.

¹⁰ U.S. Pat No. 5,580,858, issued Dec. 3, 1996, to R. M. Ippolito et al.

¹¹ M. Dubois et al., Anal. Chem., 28, (1979) 350-356.

¹² T. Mukaiyama et al., Tetrahedron Letters, 56, 5907-5908 (1968).

¹³ U.S. Pat. No. 4,137,401, issued Jan. 30, 1979, to R. Lemieux et al.

¹⁴ H. H. Westal et al., "Methods of Enzymology," 34(b), 64 (1974).

¹⁵ Svennerholm, A -M. et al., Current Microbiology, 1:19-23 (1978).

All of the above publications, and patents are herein incorporated byreference in their entirety to the same extent as if each individualpublication, patent or patent application was specifically andindividually indicated to be incorporated by reference in its entirety.

STATE OF THE ART

Toxins produced by organisms, such as bacteria, viruses, protozoa, fungiand other organisms, are known to cause a number of animal and humandiseases, including many diarrheal diseases. For example, heat-labileenterotoxin ("LT"), secreted by certain enterotoxigenic strains ofEscherichia coli, has been identified as one of the causative agents ofbacterial-induced traveller's diarrhea.¹ Additionally, cholera toxin("CT"), produced by Vibrio cholerae, has been identified as thecausative agent of the severe diarrheal disease, cholera.¹

Heat-labile enterotoxin and cholera toxin are known to bind tooligosaccharide receptors on host cells as an initial step in thepathological development of the associated disease condition.²Specifically, both LT and CT are known to bind to ganglioside G_(M1), aglycosphingolipid situated in the outer leaflet of the host cellmembrane.² G_(M1) has a characteristic pentasaccharide structure, i.e.,Gal(β1→3)GalNAc(β1→4){NeuAc(α2→3)}Gal(β1→4)Glc, on its surface whichserves as a receptor for LT and CT. LT is also known to bind to othergangliosides, such as ganglioside G_(D1b).

Additionally, many virulent organisms (e.g., bacteria, virus, fungi, andthe like) including enterovirulent organisms bind to cell surfacereceptors as part of the disease process. For example, bacteria such asVibrio cholerae and enterotoxigenic strains of Escherichia coli candirectly bind to cell surface receptors forming a colony at the point ofattachment. Such binding is detrimental because it permits expressedtoxin to immediately interact with the cell surface.

In order to ameliorate or prevent the noxious or deleterious effectscaused by toxins and organisms, it would be highly desirable to be ableto inhibit the binding of the toxin or the organism to its correspondingcell surface receptor. The present invention provides novel 1-galactosederivatives which effectively inhibit such binding.

SUMMARY OF THE INVENTION

This invention is directed to the discovery of a novel class of1-galactose derivatives which inhibit the binding of toxins, such asheat-labile enterotoxin (LT) or cholera toxin (CT), to their receptors.The compounds of this invention also inhibit binding of organisms, suchas Vibrio cholerae and enterotoxigenic strains of Escherichia coli, totheir cell surface receptors.

Accordingly, in one of its composition aspects, this invention providescompounds of formula I: ##STR1## wherein A is selected from the groupconsisting of arylene, cycloalkylene, cycloalkenylene, heteroarylene anddivalent heterocyclic;

B is selected from the group consisting of cycloalkyl, cycloalkenyl andheterocyclic;

Y is selected from the group consisting of oxygen, sulfur, --S(O)-- and--SO₂ --;

Y' is selected from the group consisting of oxygen, sulfur, --S(O)--,--SO₂ --, alkylene, substituted alkylene, and --NR¹ --, wherein R¹ isselected from the group consisting of hydrogen, alkyl and acyl; and

R^(a), R^(b), R^(c) and R^(d) are each independently selected from thegroup consisting of hydrogen; sulfate; --C(O)R², wherein R² is selectedfrom the group consisting of alkyl, alkenyl, alkaryl, alkoxyalkyl, aryl,cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic and thioalkoxyalkyl;and --P(O)(OR³)₂, wherein each R³ is independently selected from thegroup consisting of hydrogen, alkyl, alkenyl, alkaryl, alkoxyalkyl,aryl, cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic andthioalkoxyalkyl;

and pharmaceutically acceptable salts thereof.

In one preferred embodiment, the present invention is directed to theα-anomers of compounds of formula I. In another preferred embodiment,this invention is directed to the β-anomers of compounds of formula I.

In formula I above, A is preferably a cycloalkylene group having from 5to 7 carbon atoms. More preferably, A is a cycloalkylene group havingfrom 5 to 7 carbon atom wherein the cycloalkylene group is substitutedwith 1 to 3 alkyl groups. Still more preferably, A is a cyclopentylene,methylcyclopentylene, dimethylcyclopentylene, cyclohexylene,methylhexylene, dimethylcyclohexylene or cycloheptylene group. Stillmore preferably, A is a dimethylcyclopentylene group.

Preferably, B is a cycloalkyl group having from 4 to 7 carbon atoms.More preferably, B is a cycloalkyl group having 4 to 7 carbon atomswherein the cycloalkyl group is substituted with 1 to 3 alkyl groups.More preferably, B is a cyclobutyl, methylcyclobutyl,dimethylcyclobutyl, cyclopentyl, methylcyclopentyl, dimethylcyclopentyl,cyclohexyl, dimethylcyclohexyl or cycloheptyl group. Still morepreferably, B is a cyclobutyl or dimethylcyclobutyl group.

In one preferred embodiment of this invention, Y is sulfur (i.e.,--S--). In another preferred embodiment, Y is oxygen (i.e., --O--).

Preferably, Y' is --NH--.

Preferably, R^(a), R^(b), R^(c) and R^(d) are each independentlyselected from the group consisting of hydrogen and --C(O)R², where R² isalkyl. More preferably, R^(a), R^(b), R^(c) and R^(d) are each hydrogen.

Particularly preferred compounds provided by this invention include, byway of example, the following:

2,2-dimethyl-4-(cyclobutylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

2,2-dimethyl-4-(3,3-dimethylcyclobut-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

2,2-dimethyl-4-(cyclopentylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

2,2-dimethyl-4-(3-methylcyclopent-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

2,2-dimethyl-4-(3,3-dimethylcyclopent-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

2,2-dimethyl-4-(cyclohexylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

2,2-dimethyl-4-(3-methylcyclohex-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

2,2-dimethyl-4-(4-methylcyclohex-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside

and pharmaceutically acceptable salts thereof.

In another of its composition aspects, this invention provides apharmaceutical composition comprising from 1 to 99 weight percent of apharmaceutically acceptable carrier and from 1 to 99 weight percent ofat least one compound of formula I above.

In one of its method aspects, this invention is directed to a method ofameliorating conditions associated with binding of a toxin to itsreceptor in an animal which method comprises administering to saidanimal an effective amount of a pharmaceutical composition comprisingfrom 1 to 99 weight percent of a pharmaceutically acceptable carrier andfrom 1 to 99 weight percent of at least one compound of formula I above,wherein the compound of formula I inhibits the binding of the toxin toits receptor.

In preferred embodiments of this invention, the toxin in the abovemethod is heat-labile enterotoxin or cholera toxin.

In another of its method aspects, this invention is directed to a methodof ameliorating conditions associated with binding of an organism to itscell surface receptor in an animal which method comprises administeringto said animal an effective amount of a pharmaceutical compositioncomprising from 1 to 99 weight percent of a pharmaceutically acceptablecarrier and from 1 to 99 weight percent of at least one compound offormula I above, wherein the compound of formula I inhibits the bindingof the organism to its cell surface receptor.

In preferred embodiments of this invention, the organism in the abovemethod is Vibrio cholerae or an enterotoxigenic strain of Escherichiacoli.

This invention is also directed to 1-galactose derivative-containingsupports which are useful for inhibiting the binding of a toxin to itsreceptor. Supports useful for inhibiting the binding of an organism toits cell surface receptor are also provided.

Accordingly, in yet another of its composition aspects, this inventionprovides a 1-galactose derivative-containing support comprising asupport having covalently bound thereto a plurality of at least onecompound of formula I': ##STR2## wherein A is selected from the groupconsisting of arylene, cycloalkylene, cycloalkenylene, heteroarylene anddivalent heterocyclic;

B is selected from the group consisting of cycloalkyl, cycloalkenyl andheterocyclic;

Y is selected from the group consisting of oxygen, sulfur, --S(O)-- and--SO₂ --;

Y' is selected from the group consisting of oxygen, sulfur, --S(O)--,--SO₂ --, alkylene, substituted alkylene, and --NR³ --, wherein R³ isselected from the group consisting of hydrogen, alkyl and acyl; and

R^(a), R^(b), R^(c) and R^(d) are each independently selected from thegroup consisting of hydrogen; sulfate; --C(O)R', wherein R' is selectedfrom the group consisting of alkyl, alkenyl, alkaryl, alkoxyalkyl, aryl,cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic and thioalkoxyalkyl;and --P(O)(OR²)₂, wherein each R² is independently selected from thegroup consisting of hydrogen, alkyl, alkenyl, alkaryl, alkoxyalkyl,aryl, cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic andthioalkoxyalkyl;

and pharmaceutically acceptable salts thereof;

wherein one of A, B, R^(a), R^(b), R^(c) or R^(d) is covalently boundvia a linking arm to the support.

In still another of its composition aspects, this invention provides apharmaceutical composition comprising from 1 to 99 weight percent of apharmaceutically acceptable carrier and from 1 to 99 weight percent of a1-galactose derivative-containing support.

In another of its method aspects, this invention is directed to a methodof ameliorating conditions associated with binding of a toxin to itsreceptor in an animal which method comprises administering to saidanimal an effective amount of a pharmaceutical composition comprisingfrom 1 to 99 weight percent of a pharmaceutically acceptable carrier andfrom 1 to 99 weight percent of a 1-galactose derivative-containingsupport, wherein the compound of formula I' inhibits the binding of thetoxin to its receptor.

In another of its method aspects, this invention is directed to a methodof ameliorating conditions associated with binding of an organism to itscell surface receptor in an animal which method comprises administeringto said animal an effective amount of a pharmaceutical compositioncomprising from 1 to 99 weight percent of a pharmaceutically acceptablecarrier and from 1 to 99 weight percent of a 1-galactosederivative-containing support, wherein the compound of formula I'inhibits the binding of the organism to its cell surface receptor.

In a preferred embodiment of this invention, the support employed in theabove compositions and methods is a non-absorbable support. Morepreferably, the support is a non-absorbable solid support.

Preferred compounds of formula I above for use in this invention includethose set forth in formula IA below: ##STR3## wherein A, B, Y, and Y'are selected as shown in Table I below.

                  TABLE I    ______________________________________    Y     A             Y'       B    ______________________________________    --S-- 2,2-dimethylcyclopent-                        --NH--   cyclobut-1-yl          1,4-diyl    --S-- 2,2-dimethylcyclopent-                        --NH--   3,3-dimethylcyclobut-1-yl          1,4-diyl    --S-- 2,2-dimethylcyclopent-                        --NH--   cyclopent-1-yl          1,4-diyl    --S-- 2,2-dimethylcyclopent-                        --NH--   3-methylcyclopent-1-yl          1,4-diyl    --S-- 2,2-dimethylcyclopent-                        --NH--   3,3-dimethycyclopent-1-yl          1,4-diyl    --S-- 2,2-dimethylcyclopent-                        --NH--   cyclohex-1-yl          1,4-diyl    --S-- 2,2-dimethylcyclopent-                        --NH--   3-methylcyclohex-1-yl          1,4-diyl    --S-- 2,2-dimethylcyclopent-                        --NH--   4-methylcyclohex-1-yl          1,4-diyl    ______________________________________

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates a preferred reaction scheme which can be used toprepare various 1-galactose derivatives from an α,β-unsaturated cycliccarbonyl compound, i.e., cyclopent-2-en-1-one.

FIG. 2 illustrates a preferred reaction scheme which can be used toprepare various 1-galactose derivatives from an α-halocarbonyl compound,i.e, 2-chlorocyclopentan-1-one.

DETAILED DESCRIPTION OF THE INVENTION

This invention relates, in one embodiment, to compounds which inhibitthe binding of a toxin, such as heat-labile enterotoxin or choleratoxin, to its receptor either in vitro or in vivo. In anotherembodiment, the compounds of this invention inhibit binding of anorganism (e.g., bacteria, virus, fungi, and the like), such as Vibriocholerae or enterotoxigenic strains of Escherichia coli, to its cellsurface receptor. However, prior to describing this invention in furtherdetail, the following terms will first be defined.

DEFINITIONS

"Acyl" refers to the groups alkyl-C(O)--, aryl-C(O)--, andheteroaryl-C(O)-- where alkyl, aryl and heteroaryl are as definedherein.

"Acylamino" refers to the group --C(O)NRR where each R is independentlyhydrogen or alkyl.

"Acyloxy" refers to the groups alkyl-C(O)O--, aryl-C(O)O--,heteroaryl-C(O)O--, and heterocyclic-C(O)O-- where alkyl, aryl,heteroaryl and heterocyclic are as defined herein.

"Alkaryl" refers to -alkylene-aryl groups preferably having from 1 to 8carbon atoms in the alkylene moiety and from 6 to 10 carbon atoms in thearyl moiety. Such alkaryl groups are exemplified by benzyl, phenethyland the like.

"Alkoxy" refers to the group alkyl-O--. Such alkoxy groups include, byway of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy,tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, andthe like.

"Alkoxyalkyl" refers to the group -alkylene-O-alkyl which includes byway of example, methoxymethyl (CH₃ OCH₂ --), methoxyethyl (CH₃ --O--CH₂CH₂ --) and the like.

"Alkenyl" refers to alkenyl groups preferably having from 2 to 8 carbonatoms and more preferably 2 to 6 carbon atoms and having at least 1 andpreferably from 1-2 sites of alkenyl unsaturation. Such alkenyl groupsinclude ethenyl (--CH═CH₂), n-propenyl (i.e., allyl) (--CH₂ CH═CH₂),iso-propenyl (--C(CH₃)═CH₂), and the like.

"Alkyl" refers to monovalent alkyl groups preferably having from 1 to 8carbon atoms and more preferably 1 to 6 carbon atoms. This term isexemplified by groups such as methyl, ethyl, n-propyl, iso-propyl,n-butyl, iso-butyl, n-hexyl, and the like.

"Substituted alkyl" refers to a branched or straight chain alkyl groupof from 1 to 8 carbon atoms having from 1 to 3 substituents selectedfrom the group consisting of hydroxy, acyl, acylamino, acyloxy, alkoxy,alkenyl, alkynyl, amino, aminoacyl, aryl, aryloxy, carboxy,carboxyalkyl, cyano, cycloalkyl, guanidino, halo, heteroaryl,heterocyclic, nitro, thiol, thioaryloxy, thioheteroaryloxy, and thelike. Preferred substituents include hydroxy and amino.

"Alkylene" or "alkyldiyl" refers to divalent alkylene groups preferablyhaving from 1 to 8 carbon atoms and more preferably 1 to 6 carbon atoms.This term is exemplified by groups such as methylene (--CH₂ --),ethylene (--CH₂ CH₂ --), the propylene isomers (e.g., --CH₂ CH₂ CH₂ --and --CH(CH₃)CH₂ --) and the like.

"Substituted alkylene" or "substituted alkyldiyl" refers to divalentalkylene groups of from 1 to 8 carbon atoms having from 1 to 3substituents selected from the group consisting of hydroxy, acyl,acylamino, acyloxy, alkoxy, alkenyl, alkynyl, amino, aminoacyl, aryl,aryloxy, carboxy, carboxyalkyl, cyano, cycloalkyl, guanidino, halo,heteroaryl, heterocyclic, nitro, thiol, thioaryloxy, thioheteroaryloxy,and the like. Preferred substituents include hydroxy and amino.

"Alkynyl" refers to alkynyl groups preferably having from 2 to 8 carbonatoms and more preferably 2 to 6 carbon atoms and having at least 1 andpreferably from 1-2 sites of alkynyl unsaturation. Such alkynyl groupsinclude ethynyl (--C.tbd.CH), propargyl (--CH₂ C.tbd.CH) and the like.

"Amino acid" refers to any of the naturally occurring amino acids, aswell as synthetic analogs and derivatives thereof. α-Amino acidscomprise a carbon atom to which is bonded an amino group, a carboxygroup, a hydrogen atom, and a distinctive group referred to as a "sidechain". The side chains of naturally occurring amino acids are wellknown in the art and include, for example, hydrogen (e.g., as inglycine), alkyl (e.g., as in alanine, valine, leucine, isoleucine,proline), substituted alkyl (e.g., as in threonine, serine, methionine,cysteine, aspartic acid, asparagine, glutamic acid, glutamine, arginine,and lysine), alkaryl (e.g., as in phenylalanine and tryptophan),substituted arylalkyl (e.g., as in tyrosine), and heteroarylalkyl (e.g.,as in histidine). One of skill in the art will appreciate that the term"amino acid" can also include β-, γ-, δ-, and ω-amino acids, and thelike. Unnatural amino acids are also known in the art, as set forth in,for example, Williams³, Evans et al.⁴, Pu et al.⁵, Williams et al.⁶, andall references cited therein. Stereoisomers (e.g., D-amino acids) of thetwenty conventional amino acids, unnatural amino acids such asα,α-disubstituted amino acids and other unconventional amino acids mayalso be suitable components for compounds of the present invention.Examples of unconventional amino acids include: 4-hydroxyproline,3-methylhistidine, 5-hydroxylysine, and other similar amino acids andimino acids (e.g., 4-hydroxyproline).

"Aminoacyl" refers to the group --NRC(O)R where each R is independentlyhydrogen or alkyl.

"Aryl" refers to an unsaturated aromatic carbocyclic group of from 6 to14 carbon atoms having a single ring (e.g., phenyl) or multiplecondensed rings (e.g., naphthyl or anthryl). Preferred aryls includephenyl, naphthyl and the like.

Unless otherwise constrained by the definition for the aryl substituent,such aryl groups can optionally be substituted with from 1 to 3substituents selected from the group consisting of hydroxy, acyl,acyloxy, alkyl, substituted alkyl, alkoxy, alkenyl, alkynyl, amino,aminoacyl, aryl, aryloxy, carboxy, carboxyalkyl, cyano, halo, nitro,heteroaryl, trihalomethyl and the like. Preferred substituents includealkyl, alkoxy, halo, carboxy, cyano, nitro, trihalomethyl, andthioalkoxy.

"Arylene" refers to a divalent unsaturated aromatic carbocyclic group offrom 6 to 14 carbon atoms having a single ring (e.g., phenyl) ormultiple condensed rings (e.g., naphthyl or anthryl). Preferred arylenesinclude phenyl, naphthyl and the like.

Unless otherwise constrained by the definition for the arylenesubstituent, such arylene groups can optionally be substituted with from1 to 3 substituents selected from the group consisting of hydroxy, acyl,acyloxy, alkyl, substituted alkyl, alkoxy, alkenyl, alkynyl, amino,aminoacyl, aryl, aryloxy, carboxy, carboxyalkyl, cyano, halo, nitro,heteroaryl, trihalomethyl and the like. Preferred substituents includealkyl, alkoxy, halo, carboxy, cyano, nitro, trihalomethyl, andthioalkoxy.

"Aryloxy" refers to the group aryl-O-- where the aryl group is asdefined herein including optionally substituted aryl groups as alsodefined herein.

"Carboxy" refers to the group --COOH.

"Carboxyalkyl" refers to the group --C(O)O-alkyl where alkyl is asdefined herein.

"Cycloalkyl" refers to cyclic alkyl groups of from 3 to 20 carbon atoms,preferably 4 to 8 carbon atoms, having a single cyclic ring or multiplecondensed rings which can be optionally substituted with from 1 to 3substituents selected from the group consisting of hydroxy, acyl,acyloxy, alkyl, substituted alkyl, alkylene, alkoxy, alkenyl, alkynyl,amino, aminoacyl, aryl, aryloxy, carboxy, carboxyalkyl, cyano, halo,nitro, heteroaryl, trihalomethyl and the like. Preferred substituentsinclude alkyl, alkoxy, halo, carboxy, cyano, nitro, trihalomethyl, andthioalkoxy. Such cycloalkyl groups include, by way of example, singlering structures such as cyclopropyl, cyclobutyl, cyclopentyl,cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl,2-methylcyclooctyl, and the like, or multiple ring structures such asadamantanyl and the like, and spiro compounds. Examples of suitablecycloalkyl rings include single ring structures such as cyclopentane,cyclohexane, cycloheptane, cyclooctane, and the like, or multiple ringstructures such as bicyclo 2.2.1!heptane, bicyclo 3.2.1!octane, and thelike. Preferred cycloalkyl rings include cyclopentane, cyclohexane,cycloheptane and bicyclo 3.2.1!octane.

"Cycloalkylene" or "cycloalkyldiyl" refers to a divalent cyclic alkylenegroup of from 3 to 20 carbon atoms, preferably 4 to 8 carbon atoms,having a single cyclic ring or multiple condensed rings which can beoptionally substituted with from 1 to 3 substituents selected from thegroup consisting of hydroxy, acyl, acyloxy, alkyl, substituted alkyl,alkylene, alkoxy, alkenyl, alkynyl, amino, aminoacyl, aryl, aryloxy,carboxy, carboxyalkyl, cyano, halo, nitro, heteroaryl, trihalomethyl andthe like. Preferred substituents include alkyl, alkoxy, halo, carboxy,cyano, nitro, trihalomethyl, and thioalkoxy. Such cycloalkylene groupsinclude, by way of example, single ring structures such ascyclopropylene, cyclobutylene, cyclopentylene (e.g.,cyclopent-1,3-diyl), cyclooctylene, 1-methylcyclopropylene,2-methylcyclopentylene, 2-methylcyclooctylene, and the like, or multiplering structures such as adamantanylene, and the like.

"Cycloalkenyl" refers to cyclic alkenyl groups of from 4 to 20 carbonatoms, preferably 5 to 8 carbon atoms, having a single cyclic ring andat least one point of internal unsaturation. Optionally, suchcycloalkenyl groups can be substituted with from 1 to 3 substituentsselected from the group consisting of hydroxy, acyl, acyloxy, alkyl,substituted alkyl, alkylene, alkoxy, alkenyl, alkynyl, amino, aminoacyl,aryl, aryloxy, carboxy, carboxyalkyl, cyano, halo, nitro, heteroaryl,trihalomethyl and the like. Preferred substituents include alkyl,alkoxy, halo, carboxy, cyano, nitro, trihalomethyl, and thioalkoxy.Examples of cycloalkenyl groups include, cyclopentenyl, cyclohexenyl,and the like.

"Cycloalkenylene" or "cycloalkenyldiyl" refers to cyclic alkenylenegroups of from 4 to 20 carbon atoms, preferably 5 to 8 carbon atoms,having a single cyclic ring and at least one point of internalunsaturation. Optionally, such cycloalkenylene groups can be substitutedwith from 1 to 3 substituents selected from the group consisting ofhydroxy, acyl, acyloxy, alkyl, substituted alkyl, alkylene, alkoxy,alkenyl, alkynyl, amino, aminoacyl, aryl, aryloxy, carboxy,carboxyalkyl, cyano, halo, nitro, heteroaryl, trihalomethyl and thelike. Preferred substituents include alkyl, alkoxy, halo, carboxy,cyano, nitro, trihalomethyl, and thioalkoxy. Examples of cycloalkenylenegroups include, for instance, cyclopentenylene, cyclohexenylene, and thelike.

"Halo" or "halogen" refers to fluoro, chloro, bromo and iodo andpreferably is either chloro or bromo.

"Heteroaryl" refers to a monovalent aromatic group of from 2 to 8 carbonatoms and 1 to 4 heteroatoms selected from oxygen, nitrogen and sulfurwithin the ring.

Unless otherwise constrained by the definition for the heteroarylsubstituent, such heteroaryl groups can be optionally substituted with 1to 3 substituents selected from the group consisting of alkyl,substituted alkyl, alkoxy, aryl, aryloxy, halo, nitro, heteroaryl,thioalkoxy, thioaryloxy and the like. Such heteroaryl groups can have asingle ring (e.g., pyridyl or furyl) or multiple condensed rings (e.g.,indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl,pyrrolyl and furyl.

"Heteroarylene" refers to a divalent aromatic group of from 2 to 8carbon atoms and 1 to 4 heteroatoms selected from oxygen, nitrogen andsulfur within the ring.

Unless otherwise constrained by the definition for the heteroarylenesubstituent, such heteroarylene groups can be optionally substitutedwith 1 to 3 substituents selected from the group consisting of alkyl,substituted alkyl, alkoxy, aryl, aryloxy, halo, nitro, heteroaryl,thioalkoxy, thioaryloxy and the like. Such heteroarylene groups can havea single ring (e.g., pyridylene or furylene) or multiple condensed rings(e.g., indolizinylene or benzothienylene). Preferred heteroarylenesinclude pyridylene, pyrrolylene and furylene.

"Heterocycle" or "heterocyclic" refers to a monovalent saturated orunsaturated group having a single ring or multiple condensed rings, offrom 1 to 8 carbon atoms and from 1 to 4 hetero atoms selected fromnitrogen, sulfur or oxygen within the ring. For the purposes of thisapplication, the term "heterocycle" or "heterocyclic" does not includecarbohydrate rings (i.e. mono- or oligosaccharides).

Unless otherwise constrained by the definition for the heterocyclicsubstituent, such heterocyclic groups can be optionally substituted with1 to 3 substituents selected from the group consisting of alkyl,substituted alkyl, alkylene, alkoxy, aryl, aryloxy, halo, nitro,heteroaryl, thioalkoxy, thioaryloxy and the like. Such hetercyclicgroups can have a single ring (e.g., pyrrolidinyl, piperidinyl,morpholinyl or tetrahydrofuranyl) or multiple condensed rings (e.g.,indolinyl).

"Heterocyclene" or "divalent heterocyclic" refers to a divalentsaturated or unsaturated group having a single ring or multiplecondensed rings, of from 1 to 8 carbon atoms and from 1 to 4 heteroatoms selected from nitrogen, sulfur or oxygen within the ring. For thepurposes of this application, the term "heterocyclene" or "divalentheterocyclic" does not include divalent carbohydrate rings (i.e. mono-or oligosaccharides).

Unless otherwise constrained by the definition for the divalentheterocyclic substituent, such divalent heterocyclic groups can beoptionally substituted with 1 to 3 substituents selected from the groupconsisting of alkyl, substituted alkyl, alkylene, alkoxy, aryl, aryloxy,halo, nitro, heteroaryl, thioalkoxy, thioaryloxy and the like. Suchdivalent heterocyclic groups can have a single ring or multiplecondensed rings.

Examples of nitrogen heterocycles and heteroaryls include, but are notlimited to, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine,quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine,quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,phenanthridine, acridine, phenanthroline, isothiazole, phenazine,isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline,piperidine, piperazine, indoline and the like.

"Thioalkoxyalkyl" refers to the group -alkylene-S-alkyl which includesby way of example, thiomethoxymethyl (CH₃ SCH₂ --), thiomethoxyethyl(CH₃ --S--CH₂ CH₂ --) and the like.

"Thiol" refers to the group --SH.

"Thioalkoxy" refers to the group --S-alkyl wherein the alkyl group is asdefined herein.

"Thioaryloxy" refers to the group aryl-S-- wherein the aryl group is asdefined herein, including optionally substituted aryl groups as alsodefined herein.

"Thioheteroaryloxy" refers to the group heteroaryl-S-- wherein theheteroaryl group is as defined herein, including optionally substitutedheteroaryl groups as also defined herein.

The term "linking arm" refers to a chemical group or covalent bond whichoptionally covalently attaches the 1-galactose derivative to a support.Such groups typically comprise an alkylene, arylene or alkarylene groupand at least one heteroatom, preferably 2 to 6 heteroatoms. Aparticularly preferred linking arm is illustrated in the formula:

    (1-galactose derivative)-NH--(CH.sub.2).sub.m --NHC(O)NH-(support)

wherein m is an integer of from 2 to about 10. Preferably, m is 6.

The term "support" refers to an inert material or molecule to which a1-galactose derivative may be covalently bound, either directly orthrough a linking arm. When used in vivo, the solid support will bebiocompatible and pharmaceutically acceptable. Preferably, the supportis a non-absorbable support, i.e., when administered orally, the supportpasses unaffected through the gut without being absorbed into thecirculatory system and is essentially completely eliminated from thebody. More preferably, the support is a non-absorbable solid support.Typically, the support will contain a plurality of attachment sites forthe 1-galactose derivative, i.e., the support is an oligovalent orpolyvalent carrier. Suitable supports range, by way of illustration,from low molecular weight molecules, such 1,3,5-benzenetricarboxylicacid (trimesic acid), to organic and inorganic polymers,polysaccharides, polypeptides, glasses, silicates or minerals.

The term "solid support" refers to an inert, non-absorbable solidmaterial to which a 1-galactose derivative may be covalently bound,preferably via a linking arm. When used in vivo, the solid support willbe biocompatible and pharmaceutically acceptable. Suitable solidsupports include, by way of example only, silica, including syntheticsilicates, such as porous glass; biogenic silicates, such asdiatomaceous earth; hydrogels; silicate-containing minerals, such askaolinite; synthetic polymers, such as polystyrene, polypropylene, etc.;polysaccharides such as dextrans, celluloses (CMC), alginates, chitins,chitosans and cyclodextrins; and the like.

Preferred solid support materials for use in this invention are silicasupports which have been silylaminated with aω-aminoalkyltrialkoxysilane using conventional procedures. Suitableω-aminoalkyltrialkoxysilanes include, for example,3-aminopropyltriethoxysilane, 4-aminobutyltriethoxysilane and the like.A particularly preferred silica for use in such silylamination reactionsis silica sold commercially under the tradename Chromosorb P™ byManville Corp., Denver, Colo.

The term "toxin" refers to a compound produced by an organism whichcauses or initiates the development of a noxious, poisonous ordeleterious effect in a host presented with the toxin. Such deleteriousconditions may include fever, nausea, diarrhea, weight loss, neurologicdisorders, renal disorders, hemorrhage, and the like. As used herein,the term "toxin" includes bacterial toxins, such as cholera toxin,heatliable and heat-stable toxins of E. coli, toxins A and B ofClostridium difficile, aerolysins, hemolysins, and the like; toxinsproduced by protozoa, such as Giardia; toxins produced by fungi; and thelike. Included within this term are exotoxins, i.e., toxins secreted byan organism as an extracellular product, and enterotoxins, i.e., toxinspresent in the gut of an organism.

The terms "heat-labile enterotoxin" or "LT" refer to an enterotoxin ofenterotoxigenic E. coli which initiates traveller's diarrhea and relatedconditions. This toxin has a lectin-like activity.

The term "traveller's diarrhea" refers to diarrhea of sudden onset,often accompanied by abdominal cramps, vomiting and fever that occurssporadically in traveller's, usually during the first week of a trip.This diarrhea is most commonly caused by enterotoxigenic E. coli.

The term "cholera" refers to an acute epidemic infectious disease causedby Vibrilo cholerae, wherein a soluble toxin elaborated in theintestinal tract by the Vibrio alters the permeability of the mucosa,causing a profuse watery diarrhea, extreme loss of fluid andelectrolytes, and a state of dehydration and collapse, but no grossmorphologic change in the intestinal mucosa.

The terms "cholera toxin" or "CT" refer to an enterotoxin of V. choleraewhich initiates cholera and related conditions. This toxin has alectin-like activity.

The phrase "inhibit(s) the binding of a toxin to its receptor" meansthat a compound inhibits the binding of a toxin to its receptor by atleast 20%. For example, useful binding inhibition assays may measureinhibition of binding to ganglioside G_(D1b) or ganglioside G_(M1),neutralization of cytotoxic activity, or the like. Such binding isreported herein as percent toxin activity remaining so that thosecompounds which result in about 80% or less toxin activity remainingunder the bioassay conditions disclosed herein are deemed to inhibit thebinding of a toxin to its receptor.

The phrase "inhibit(s) the binding of heat-labile enterotoxin (LT)and/or cholera toxin (CT) to an LT and/or CT receptor" means that acompound inhibits the binding of LT and/or CT to an LT and/or CTreceptor by at least 20%.

The phrase "inhibit(s) the binding of an organism to its cell surfacereceptor" means that a compound inhibits the binding of an organism,such as a bacterium, a virus, a protozoan, a fungus, and the like, toits cell surface receptor. For example, for organisms such as Vibrocholera or enterotoxigenic strains of E. coli, a compound is said toinhibit binding of an organism to a cell surface receptor if it reducesbinding of a bacterial surface adhesion antigen, such as CFA I pili, byat least 10%.

The term "pharmaceutically acceptable salt" refers to pharmaceuticallyacceptable salts of a compound of formula I which salts are derived froma variety of organic and inorganic counter ions well known in the artand include, by way of example only, sodium, potassium, calcium,magnesium, ammonium, tetraalkylammonium, and the like; and when themolecule contains a basic functionality, salts of organic or inorganicacids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate,maleate, oxalate and the like.

For purpose of this application, all sugars are referenced usingconventional three letter nomenclature. All sugars are assumed to be inthe D-form unless otherwise noted, except for fucose, which is in theL-form. Further, all sugars are in the pyranose form.

When chiral centers are found in the 1-galactose derivatives of thisinvention other than the chiral centers of the galactose moiety, thisinvention encompasses all possible stereoisomers, i.e., enantiomers ordiastereomers. For example, when A is a cycloalkylene group, the carbonatoms to which Y and Y' are attached may have an R,R or R,S or S,R orS,S configuration. Similarly, B is a cycloalkyl group, the carbon atomto which Y' are attached may have an R or S configuration.

General Synthetic Procedures

The 1-galactose derivatives of this invention may be prepared by thefollowing general methods and procedures. It should be appreciated thatwhere typical or preferred process conditions (e.g., reactiontemperatures, times, mole ratios of reactants, solvents, pressures,etc.) are given, other process conditions may also be used unlessotherwise stated. Optimum reaction conditions may vary with theparticular reactants or solvents used, but such conditions can bedetermined by one skilled in the art by routine optimization procedures.

The 1-galactose derivatives of this invention where Y is sulfur,--S(O)--, or S(O)₂ and A is cycloalkylene, cycloalkenylene or divalentheterocyclic (i.e., A is non-aromatic) are typically prepared byreaction of a 2,3,4,6-tetra-O-protected 1-thiogalactose intermediatewith a cyclic α,β-unsaturated carbonyl compound or a cyclicα-halocarbonyl compound. The resulting carbonyl-containing intermediateis then reduced or reductively aminated to give an alcohol or an aminecompound. These alcohol or amine compounds are then further reacted viareductive alkylation or by conversion to a leaving group anddisplacement to afford amines, ethers or thioethers and the like. Thecarbonyl-containing intermediate can also be reductively aminated toafford amines. Such reactions are well known to those of ordinary skillin the art and can be accomplished using art recognized procedures.

The cyclic α,β-unsaturated carbonyl compounds suitable for use inpreparing the 1-galactose derivatives of this invention are well knownin the art. Such compound are either commercially available or can beprepared from commercially available materials using art recognizedprocedures.

Preferred cyclic α,β-unsaturated carbonyl compounds for use in thisinvention include, by way of example, cyclopent-2-en-1-one,4,4-dimethylcyclopent-2-en-1-one, cyclohex-2-en-1-one,4,4-dimethylcyclohex-2-en-1-one, 6,6-dimethylcyclohex-2-en-1one andcyclohept-en-1-one.

The cyclic α-halocarbonyl compounds employed in preparing the1-galactose derivatives of this invention are also well known in theart. Such compounds are either commercially available or can be preparedfrom commercially available materials using art recognized procedures.Preferred cyclic α-halocarbonyl compounds for use in this inventioninclude, by way of example, 2-chlorocyclopentanone and2-chlorocyclohexanone. Alternatively, cyclic carbonyl compounds having aleaving group other than a halogen in the α-position may be employed.Suitable leaving groups include, by way of illustration, varioussulfonic ester groups, such as tosylate, mesylate, brosylate andnosylate groups and the like, and fluorinated sulfonic ester groups,such as triflate, nonaflate and tresylate groups and the like.

The synthesis of various 1-galactose derivatives from either a cyclic α,β-unsaturated carbonyl compound or a cyclic α-halocarbonyl compound isillustrated in FIGS. 1 and 2, respectively. FIG. 1 illustrates thesynthesis of various 1-galactose derivatives from cyclopent-2-en-1-one.FIG. 2 illustrates the synthesis of various 1galactose from2-chlorocyclopentan-1-one. It will be readily apparent to those ofordinary skill in the art that the synthetic procedure illustrated inFIGS. 1 and 2 and the following reaction conditions can be modified byselecting the appropriate starting materials and reagents to allow thepreparation of other 1-galactose derivatives of this invention.

As shown in FIG. 1, D-galactose is perlauroylated by contactingD-galactose with at least 5 equivalents, and preferably 10 equivalents,of lauroyl chloride. This reaction is generally conducted in an inertdiluent, such pentane, hexane, dichloromethane and the like, using atertiary amine such as pyridine or triethylamine to neutralize thehydrochloric acid generated during the reaction. Preferably, a catalyticamount of 4-(N,N-dimethylamino)pyridine is added to the reaction mixtureto facilitate this reaction. Typically, this reaction is conducted at atemperature of from about -78° C. to about 30° C. for about 0.5 to about96 hours to afford 1,2,3,4,6-penta-O-lauroyl-α-D-galactopyranose, 1, inapproximately 70% yield from D-galactose.

Compound 1 is then converted into1-S-acetyl-2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranose, 2, byreaction of 1 with an excess of thiolacetic acid. In one embodiment,this reaction is conducted in the presence of an excess of borontrifluoride etherate, preferably using about 15 to 20 equivalents ofboron trifluoride etherate based on 1, in an inert diluent, such asdichloromethane and the like. Typically, this reaction is conductedinitially at about 0° C. and then at about 20° C. to about 30° C. forabout 0.5 to about 48 hours.

In another embodiment, compound 2 can be prepared from 1 by contacting 1with at least one equivalent, preferably 1 to 1.2 equivalents, ofbenzylamine to selectively remove the 1-lauroyl group. This reaction istypically conducted at about 25° C. to about 30° C. for about 1 to about96 hours to provide for 2,3,4,6-tetra-O-lauroyl-(α,β)-galactopyranoside.This intermediate is then converted into anO-(2,3,4,6-tetra-O-lauroyl-(α,β)-galactopyranosyl)trichloroacetimidateintermediate by contacting the tetralauroyl compound with an excess oftrichloroacetonitrile, preferably about 10 equivalents, and about 0.8 toabout 1.0 equivalents, of 1,8-diazabicyclo 5.4.0!undec-7-ene (DBU) in aninert diluent, such as dichloromethane. The resultingO-trichloroacetidate intermediate is then contacted with an excess ofthiolacetic acid in an inert diluent, such as dichloromethane, at about25° C. to about 30° C. for about 1 to about 96 hours to provide for1-S-acetyl-2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranose, 2.

In still another embodiment, compound 2 can be prepared by contactingcompound 1 with about 1.5 to about 2.0 equivalents of thiolacetic acidand about 0.5 equivalents of trimethylsilyl trifluoromethanesulfonatebased on 1 in an inert diluent, such as dichloromethane and the like.Typically, this reaction is conducted initially at about 0C and then atabout 20° C. to about 30° C. for about 0.5 to about 72 hours. Thismethod is especially preferred since it provides the highest yield ofcompound 2 and produces no detectable traces of the correspondingα-isomer.

If desired, however, the α-isomer, i.e.,1-S-acetyl-2,3,4,6-tetra-O-lauroyl-1-thio-α-D-galactopyranose, can bereadily prepared by contacting compound 1 with an excess, preferablyabout 20 equivalents, of thioacetic acid in the presence of about 1.0 to1.1 equivalents of tin (IV) chloride in an inert diluent, such toluene,at ambient temperature for about 0.5 to about 2 hours. Alternatively,treatment of compound 1 with an excess, preferably about 3 to about 6equivalents, of thioacetic acid in the presence of about 2.0 to 3.0equivalents of trimethylsilyl trifluoromethanesulfonate in an inertdiluent, such dichloromethane, at ambient temperature for about 12 toabout 48 hours affords1-S-acetyl-2,3,4,6-tetra-O-lauroyl-1-thio-α-D-galactopyranose.

The Michael addition of compound 2 to cyclopent-2-en-1-one then affords3-oxocyclopentan-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-α-D-galactopyranoside, 3. This reactionis typically conducted by contacting 2 with at least one equivalent,preferably 1.0 to 1.2 equivalents, of cyclopent-2-en-1-one in thepresence of a molar excess of a dialkylamine, such as diethylamine.

Without being limited by any theory, it is believed that thedialkylamine first reacts with the thioacetyl of compound 2 therebyforming in situ the thiol derivative of compound 2 which then reactsunder basic conditions generated by the dialkylamine with a Michaeladduct.

Typically, this reaction is conducted in an inert diluent, such asdichloromethane, at a temperature of from about -40° C. to about 50° C.for about 1 to about 6 hours.

The carbonyl group of compound 3 can then reduced using a reducing agentto provide for 3-hydroxycyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 4. Preferably,this reduction is conducted by contacting 3 with sodium borohydride,preferably about 1.2 to about 2.0 equivalents of sodium borohydridebased on 3. Generally, this reaction is conducted in an inert diluent,such as tetrahydrofuran, isopropanol and mixture thereof, at atemperature of about 25° C. to about 30° C. for about 0.5 to about 3.0hours. The resulting alcohol, 4, is readily purified by solid-phaseextraction on C18 silica gel using pentane as an eluent.

The hydroxyl group of alcohol derivative 4 can then be converted into aleaving group, such as the mesylate, tosylate, etc., and displaced withvarious nucleophiles. For example, treatment of 4 with an excess,preferably about 1.1 to about 1.5 equivalents, of methanesulfonylchloride in pyridine and an inert diluent, such as THF, affords3-methanesulfonylcyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 5.

The mesylate group of 5 can then be displaced with, for example, a thiolcompound of the formula HS-B (where B is as defined above) under basicconditions to provide a thioether. For example, treatment of 5 withabout 1.0 to about 1.5 equivalents of cyclopentanethiol in the presenceof a suitable base, such as DBU, in an inert diluent, such as toluene,affords 3-(thiocyclopentoxy)cyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 6.

The lauroyl groups can then be removed from compound 6 by contacting 6with an excess of sodium methoxide in methanol and an inert diluent,such as dichloromethane, at about 25° C. to about 30° C. for about 1 toabout 24 hours. Neutralization of the reaction mixture with AmberliteIR-50S (H⁺) resin then provides for 3-(thiocyclopentoxy)cyclopent-1-yl1-thio-β-galactopyranoside, 7.

Alternatively, the mesylate group of compound 5 can be displaced with analkali or alkaline earth metal alkoxides to afford ethers. Typically,this reaction is conducted by contacting an alcohol of the formula HO-B(where B is as defined above), such as cyclopentanol, with a strongbase, such as sodium hydride, potassium hydride, calcium hydride and thelike, in an inert diluent, such as tetrahydrofuran, toluene and thelike, under substantially anhydrous conditions at a temperature in therange of from about -10° C. to about 120° C. for about 0.25 to about 3hours.

The resulting alkali or alkaline earth metal alkoxide is generally notisolated, but is reacted in situ with the mesylate compound 5 toprovide, after neutralization, an ether compound, e.g.,3-(cyclopentoxy)cyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 8. This reactionis typically conducted in a substantially anhydrous inert diluent at atemperature of from about 0° C. to about 100° C. for about 2 to about120 hours. Suitable diluents for this reaction include, tetrahydrofuran,toluene and the like.

Removal of the lauroyl from compound 8 using excess sodium methoxide inmethanol, followed by neutralization of the reaction mixture withAmberlite IR-50S (H⁺) resin then affords 3-(cyclopentoxy)cyclopent-1-yl1-thio-β-galactopyranoside, 9.

The mesylate group of compound 5 can also be displaced with sodium azideto provide, after reduction of the azido group, a primary aminecompound, e.g., 3-aminocyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 10. The azidedisplacement reaction is typically conducted by contacting the mesylatecompound 5 with an excess, preferably about 5 to about 50 equivalents ofsodium azide in an inert diluent, such as N,N-dimethylformamide, THF,and mixtures thereof, at a temperature of from about 50° C. to about100° C. for about 1 to about 6 hours. Preferably, a crown ether, such as18-crown-6, is added to the reaction mixture to promote the displacementreaction.

The azido intermediate can then be reduced with a reducing agent toafford the corresponding primary amine, i.e., compound 10. Preferably,this reaction is conducted by contacting the azido compound with about1.0 to about 1.1 equivalents of sodium borohydride and about 2.0 toabout 2.2 equivalents of nickel chloride (NiCl₂) in an inert diluent,such as ethanol, isopropanol, or mixtures thereof, at a temperature offrom about 0° C. to about 50° C. for about 0.5 to about 6 hours.

Alternatively, compound 3 can be reductively aminated to provide forcompound 10 directly. In one embodiment of this reaction, compound 3 iscontacted with an excess of ammonium acetate and at least one equivalentof sodium cyanoborohydride based on 3. This reaction is typicallyconducted in an inert diluent, such as methanol, tetrahydrofuran andmixtures thereof, at a temperature of about 25° C. to about 30° C. forabout 1 to about 72 hours.

In another preferred embodiment, the reductive amination reaction isaccomplished by contacting compound 3 with an excess of ammonium acetateand an excess of trimethyl orthoformate based on 3, in an inert diluent,such as 1,2dichloroethane at a temperature of about 25° C. to about 30°C. for about 12 to about 72 hours to form an imine intermediate. Theimine intermediate is generally not isolated but is contacted in situwith an excess of sodium borohydride, preferably about 1.2 to about 1.5equivalents of sodium borohydride, based on 3. The resulting aminocompound 10 is then readily purified by solid-phase extraction on C18silica gel using pentane as an eluent.

The primary amine group of compound 10 can then be reductively alkylatedusing a cyclic ketone, such as pentan-1-one, to afford a secondaryamine, e.g., 3-(cyclopentylamino)cyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 11. Typically,this reaction is conducted by contacting the primary amine with anexcess, preferably about 2 to about 500 equivalents of an aldehyde or aketone in the presence of at least one equivalent, preferably about 1.0to about 10 equivalents, of a reducing agent, such as sodiumtriacetoxyborohydride. This reaction is typically conducted in an inertdiluent, such as dichloromethane, methanol, or mixtures thereof, at atemperature of about 0° C. to about 50° C. for about 10 to about 48hours.

As shown in FIG. 1, compound 3 can also be reductively aminated with anamine compound of the formula H₂ N--B (where B is as defined above),such as cyclopentylamine, to provide a secondary amine compound, e.g.,compound 11. Specifically, compound 3 is contacted with a molar excessof the amine compound, preferably with 10 equivalents based on 3, in thepresence of at least one molar equivalent, preferably about 1.0 to about1.2 equivalents, of sodium cyanoborohydride. Typically, this reaction isconducted in an essentially anhydrous inert diluent, such asacetonitrile, at a temperature of about 25° C. to about 30° C. for about1 to about 72 hours. The resulting secondary amine 11 is readilypurified by solid-phase extraction on C18 silica gel using pentane asthe eluent.

The lauroyl groups of compound 11 are then removed by contacting thelauroyl-protected compound with an excess of sodium methoxide inmethanol and an inert diluent, such as dichloromethane, at about 25° C.to about 30° C. for about 1 to about 24 hours. Neutralization of thereaction mixture with Amberlite IR-50S (H⁺) resin then provides3-(cyclopentylamino)cyclopent-1-yl 1-thio-β-D-galactopyranoside, 12.

Additionally, compound 3 can be reacted with an ylide of a phosphoniumsalt (i.e., a Wittig reagent), such (cyclopentylmethyl)(triphenyl)phosphonium bromide, to afford, after hydrogenation of theresulting olefin, 3-(cyclopentylmethyl)cyclopent-1-yl2,3,4,6-tetra-O-lauroyl-l-thio-β-D-galactopyranoside, 13. This reactionis typically conducted by first contacting the phosphonium salt with aslight excess, preferably about 1. 1 to about 1.2 equivalents, of astrong base, such as n-butyl lithium, in an inert diluent, such asdiethyl ether, THF and the like, at a temperature of from about -78° C.to about 0° C. for 0.5 to 6 hours to form the ylide. Typically, theylide is not isolated but is reacted in situ with a carbonyl compound,such as 3, to afford an olefin. The resulting olefin can then readilyhydrogenated by treatment with hydrogen in the presence of a catalyst,such as Pd/C, in an inert diluent, such as ethanol, at a temperature offrom about 0° C. to about 50° C. for about 1 to 48 hours to providecompound 13. Removal of the lauroyl from compound 13 using excess sodiummethoxide in methanol, followed by neutralization of the reactionmixture with Amberlite IR-50S (H⁺) resin then affords3-(cyclopentylmethyl)cyclopent-1-yl 1-thio-O-galactopyranoside, 14.

As noted above, FIG. 2 illustrates the synthesis of various1-thiogalactose derivatives using a cyclic α-halocarbonyl carbonylcompound, i.e., 2-chloropentan-1-one. As shown in FIG. 2,1-S-acetyl-2,3,4,6-tetra-O-lauroyl-l-thio-β-D-galactopyranose, 2,prepared as described above, reacts with 2-chlorocyclopentan-1one togive 2-oxocyclopentan-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 15. This reactionis typically conducted by contacting 2 with at least one equivalent,preferably 1.0 to 1.2 equivalents, of 2-chlorocyclopentan-1-one in thepresence of an excess of a dialkylamine, such as diethylamine.Typically, this reaction is conducted in an inert diluent, such asdichloromethane, at a temperature of from about -40° C. to about 50° C.for about 1 to about 6 hours to afford compound 15.

Compound 15 can then be reacted using the same reagents and conditionsdescribed above for compound 3 to afford various 1-galactosederivatives, such as compounds 24 and 26, as illustrated in FIG. 2.

As shown in FIG. 2, ether derivative 21 is preferably prepared byalkylation of the hydroxyl compound 16. Typically, this reaction isconducted by contacting compound 16 with a strong base, such as sodiumhydride, potassium hydride, calcium hydride and the like, in an inertdiluent, such as tetrahydrofuran, toluene and the like, undersubstantially anhydrous conditions at a temperature in the range of fromabout 10° C. to about 120° C. for about 0.25 to about 3 hours. Theresulting alkali or alkaline earth metal alkoxide is generally notisolated, but is reacted in situ with, for example, a mesylate of theformula MsO--B (wherein B is as defined herein), such as1-(methanesulfonyloxy)cyclopentane, to provide after neutralization theether compound, e.g., 2-(cyclopentoxy)cyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 20. This reactionis typically conducted in a substantially anhydrous inert diluent at atemperature of from about 0° C. to about 100° C. for about 2 to about120 hours. Suitable diluents for this reaction include, tetrahydrofuran,toluene and the like.

Removal of the lauroyl from compound 20 using excess sodium methoxide inmethanol, followed by neutralization of the reaction mixture withAmberlite IR-50S (H⁺) resin then affords 2-(cyclopentoxy)cyclopent-1-yl1-thio-β-galactopyranoside, 21.

Similarly, the thioether compound 19 is prepared by first forming thethioketone deriviative of compound 15 by treatment of 15 with Lawesson'sreagent under conventional reaction conditions. The thioketone is thenreduced using a metal hydride reducing agent, such as sodiumborohydride, to afford 2-thiocyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 17. Compound 17can then be readily alkylated with, for example, a mesylate of theformula MsO--B (wherein B is as defined herein), such as1-(methanesulfonyloxy)cyclopentane, to provide after neutralization thethioether compound, e.g., 2-(thiocyclopentoxy)cyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside, 18. Removal of thelauroyl from compound 18 using excess sodium methoxide in methanol,followed by neutralization of the reaction mixture with Amberlite IR-50S(H⁺) resin then affords 2-(thiocyclopentoxy)cyclopent-1-yl1-thio-β-galactopyranoside, 19.

Optionally, the 1-galactose derivatives of formula I wherein Y is asulfide linking group (--S--) can be oxidized using conventionalreagents and conditions to provide the corresponding sulfoxide(Y=--S(O)--) and sulfone (Y=--SO₂ --) derivatives. Suitable reagents foroxidizing a sulfide compound to a sulfoxide include, by way of example,hydrogen peroxide, peracids such as 3-chloroperoxybenzoic acid (MCPBA),sodium periodate, sodium chlorite, sodium hypochlorite, calciumhypochlorite, tert-butyl hypochlorite and the like. Chiral oxidizingreagents (optically active reagents) may also be employed to providechiral sulfoxides. Such optically active reagents are well known in theart and include, for example, the reagents described in Kagen et al.⁷and references cited therein.

The oxidation reaction is typically conducted by contacting the1-galactose derivative with about 0.95 to about 1.1 equivalents of theoxidizing reagent in an inert diluent, such as dichloromethane, at atemperature ranging from about 0° C. to about 50° C. for about 1 toabout 48 hours. The resulting sulfoxide can then be further oxidized tothe corresponding sulfone by contacting the sulfoxide with at least oneadditional equivalent of an oxidizing reagent, such as hydrogenperoxide, MCPBA, potassium permanganate and the like. Alternatively, thesulfone can be prepared directly by contacting the sulfide with at leasttwo equivalents, and preferably an excess, of the oxidizing reagent.

In a similar manner, the 1-galactose derivatives of formula I where Y'is sulfur can be oxidized to afford the corresponding sulfoxide(Y'=--S(O)--) and sulfone (Y'=--SO₂ --) derivatives.

The 1-galactose derivatives of this invention where Y is oxygen and A iscycloalkylene, cycloalkenylene or divalent heterocyclic (i.e., A isnon-aromatic) are typically prepared by reacting a2,3,4,6-tetra-O-protected 1-chloro, 1-bromo, or trichloroimidategalactose intermediate with a cyclic hydroxy carbonyl compound. Usingthe conditions and procedures described herein, the resultingcarbonyl-containing intermediate is then reduced or reductively aminatedto give an alcohol or an amine compound. The alcohol or amine compoundsare then further reacted via reductive alkylation or by conversion to aleaving group and displacement to afford amines, ethers or thioethersand the like. The carbonyl-containing intermediate can also bereductively aminated to afford amines. Such reactions are well known tothose of ordinary skill in the art and can be accomplished using artrecognized procedures.

By way of example, O-(2,3,4,6-tetra-O-benzoyl-β-galactopyranosyl)trichloroacetimidate can be coupled to a cyclic hydroxy carbonylcompound, such as 3-hydroxycycloheptan-1-one, using conventionalcoupling conditions and reagents to afford 3-oxocycloheptan-1-yl2,3,4,6-tetra-β-benzoyl-β-D-galactopyranose. Typically, this couplingreaction is conducted by contacting the trichloroacetimidateintermediate with from about 1.0 to 2.0 equivalents of the cyclichydroxy carbonyl compound in the presence of an excess of trimethylsilyltrifluoromethanesulfonate. The reaction is typically conducted at atemperature ranging from about 0° C. to about 50° C. in a suitableanhydrous diluent, such as diethyl ether and the like.

The cyclic hydroxy carbonyl compounds suitable for use in this reactionare either commercially available or can be prepared from commerciallyavailable materials using art recognized procedures. For example, cyclichydroxy carbonyl compounds can be readily prepared from cyclicα,β-unsaturated carbonyl compounds by treatment of the cyclicα,β-unsaturated carbonyl compound with sodium hydroxide and hydrogenperoxide, followed by treatment of the resulting intermediate withacetic acid and sodium iodide in acetone. Formation of such compounds isfurther described in E. Hasegawa et al.⁸ and H. Paulsen et al.⁹

The carbonyl-containing intermediate resulting from the couplingreaction can then be reacted using the same reagents and conditionsdescribed above for compound 3 to afford various 1-galactosederivatives.

In a similar manner, the 1-galactose derivatives of formula I where Y isoxygen, sulfur, --S(O)-- or --S(O)₂, and A is aryl or heteroaryl (i.e.,where A is aromatic) can be prepared by coupling a hydroxy aryl orheteroaryl compound of the formula: HO--A--Y'--B or a thiol aryl orheteroaryl compound of the formula: HS--A--Y'--B (where, in eitherformula, A is aryl or heteroaryl, and B and Y' are as defined above)with a 2,3,4,6-tetra-O-protected 1-chloro, 1-bromo, or trichloroimidategalactose intermediate using conventional coupling procedures andreagents. The hydroxy or thiol aryl/heteroaryl compounds employed inthis reaction are commercially available or can be prepared fromcommercially available materials using art recognized procedures.

If desired, the hydroxyl groups of the galactose moiety may be readilyacylated, sulfonylated or phosphorylated using art recognized proceduresand reagents to provide compounds of formula I wherein at least one ofR^(a), R^(b), R^(c), and R^(d) is O--SO₂ --OH, --C(O)R¹⁰ or--P(O)(OR¹¹)₂ or pharmaceutically acceptable salts thereof, where R¹⁰and R¹¹ are as defined above. Such acylation reactions may occur as aninitial step of the synthesis (i.e., using an acyl halide, such aslauroyl chloride, as described above) or as a post-synthetictransformation of compounds of formula I where R^(a), R^(b), R^(c), andR^(d) are each hydrogen using, for example, acyl halides, anhydrides,halophosphates, sulfur trioxide, and the like.

For example, a de-blocked hydroxyl group can be sulfonylated by treatingthe hydroxy-containing compound with an excess, preferably about 1.1 toabout 1.2 equivalents, of a pyridine:sulfur trioxide complex in an inertdiluent, such as N,N-dimethylformamide, at ambient temperature for about1 to about 24 hours. Typically, the resulting sulfate (i.e., --O--SO₂--OH) is isolated as its salt by treatment with, for example, a Na⁺resin in an inert diluent, such as methanol. Further reaction conditionssuitable for forming sulfates and phosphates can be found, for example,in U.S. Pat. No. 5,580,858¹⁰.

In another embodiment of this invention, the 1-galactose derivatives ofthis invention can be attached to a support, preferably a solid support,either through the galactose moiety or through the A or B portions ofthe molecule. Methods for attaching compounds to supports throughvarious functional groups are well known in the art and any of theseknown methods may be employed to covalently attach the 1-galactosederivatives of this invention to a support.

By way of example, a 1-galactose derivative of formula I wherein A or Bcontains a carboxylic acid moiety can be covalently attached to anaminated solid support using conventional coupling procedures andreagents. Typically, such a coupling reaction will be conducted usingwell-known coupling reagents such as carbodiimides, BOP reagent(benzotriazol-1-yloxy-tris(dimethylamino)phosphoniumhexafluorophosphonate) and the like. Suitable carbodiimides include, byway of example, dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide,1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and the like.Preferably, a well-known coupling promoter, such asN-hydroxysuccinimide, 1-hydroxybenzotriazole and the like, is alsoemployed in the reaction mixture to facilitate the coupling reaction.

The coupling reaction is typically conducted by contacting the solidsupport with an excess, preferably about 1.1 to about 10 or moreequivalents, of the 1galactose derivative (based on the number ofequivalents of amino groups present on the solid support) and at leastone equivalent, preferably about 1.5 to about 3.0 equivalents, of thecoupling reagent (based on the 1-galactose derivative) in an inertdiluent, such N,N-dimethylformamide and the like. If desired, least oneequivalent, preferably about 1.5 to about 3.0 equivalents (based on the1-galactose derivative), of a coupling promoter such as1-hydroxybenzotriazole may also be used in the reaction. Generally, thecoupling reaction is conducted at a temperature ranging from about 0° C.to about 50° C. for about 24 to about 100 hours. Upon completion of thereaction, the solid support is preferably contacted with excess aceticanhydride in methanol at a temperature ranging from about 0° C. to about40° C. for about 12 to about 24 hours to cap any unreacted amino groupspresent on the solid support. The yield of incorporation of the1-thiogalactose derivative onto the solid support can be determinedusing well-established procedures such as those described, for example,by M. Dubois et al.¹¹.

The 1-galactose derivatives of this invention can also be prepared on asolid support via solid-phase synthesis techniques. Typically, suchsolid-phase techniques involve first covalently attaching a 1-galactosecompound through a hydroxyl group on the galactose moiety to a solidsupport using conventional procedures and reagents. The covalently-bound1-galactose compound is then reacted using the procedures describedabove with a cyclic α,β-unsaturated carbonyl compound or a cyclicα-halocarbonyl compound. The resulting carbonyl-containing intermediateis then reduced or reductively aminated to give an alcohol or an aminecompound which can be further derivatized as described herein.

By way of example, 1-dithioethyl-β-D-galactopyranoside is readilyattached to a trityl chloride resin having about 0.80 to about 1.00mmol/g of active chlorine by contacting the resin with about 0.75 toabout 2.0 equivalents of 1-dithioethyl-β-D-galactopyranoside in pyridinecontaining a catalytic amount of 4-(N,Ndimethylamino)pyridine at atemperature ranging from about 25° C. to about 100° C. for about 12 to48 hours. A free thiol group at the 1-position of the covalently boundgalactose is then generated by treating the resin with dithiothreitol(Cleland's reagent) and triethylamine in an inert diluent, such asmethanol, for about 6 to 24 hours at ambient temperature. The resulting1-thio-β-D-galactopyranoside is then reacted as described above toafford a 1-thiogalactose derivative of formula I covalently attached tothe solid support resin. If desired, the 1-thiogalactose derivative canbe cleaved from the solid support resin by contacting the resin with anexcess of trifluoroacetic acid and triisopropylsilane in an inertdiluent, such as dichloromethane, at ambient temperature.

Utility

In one embodiment, the compounds of this invention are useful inblocking binding of a toxin, such as heat-labile enterotoxin or choleratoxin, to its receptor either in vitro or in vivo. In anotherembodiment, the compounds of this invention inhibit binding of anorganisms (e.g., bacteria, virus, fungi, and the like), such as Vibriocholerae and enterotoxigenic strains of Escherichia coli, to its cellsurface receptor.

Accordingly, the compounds of this invention can be used to ameliorateconditions associated with infection by an organism, includinggastrointestinal infections caused by enterovirulent organisms, such asVibrio cholerae or enterotoxigenic strains of Escherichia coli,including, by way of example, diarrhea, intestinal bleeding, abdominalpain, and the like.

When used in treating or ameliorating such conditions, the compounds ofthis invention are typically delivered to a patient in need of suchtreatment by a pharmaceutical composition comprising a pharmaceuticallyacceptable diluent and an effective amount of at least one compound ofthis invention. The amount of compound administered to the patient willvary depending upon what compound and/or composition is beingadministered, the purpose of the administration, such as prophylaxis ortherapy, the state of the patient, the manner of administration, and thelike. In therapeutic applications, compositions are administered to apatient already suffering from an infection, such as gastrointestinalinfections associated with, for example, Vibrio cholerae orenterotoxigenic strains of Escherichia coli, in an amount sufficient toat least partially arrest further onset of the symptoms of the diseaseand its complications. An amount adequate to accomplish this is definedas "therapeutically effective dose." Amounts effective for this use willdepend on the judgment of the attending clinician depending upon factorssuch as the degree or severity of the infection in the patient, the age,weight and general condition of the patient, and the like. Preferably,for use as therapeutics, the compounds described herein are administeredat dosages ranging from about 0.1 to about 10 mg/kg/day.

Such pharmaceutical compositions may contain more than one compound ofthe present invention. For example, they may contain one compound offormula I which is highly effective at inhibiting the binding of LT anda different compound of formula I which is highly effective atinhibiting the binding of enterotoxigenic E. coli to cell surfacereceptors.

When a support having a compound of formula I' covalently attached isused for treating or ameliorating conditions associated withgastrointestinal infections, supports which are non-toxic, resistant tomechanical and chemical deposition are preferred. Those supports whichpass unaffected through the gut and which are completely and rapidlyeliminated following oral administration are most preferred, since suchsupports provide for rapid clearance of the toxin and/or pathogen fromthe subject.

As noted above, the compounds administered to a patient are in the formof pharmaceutical compositions described above which can be administeredby a variety of routes including oral, rectal, transdermal,subcutaneous, intravenous, intramuscular, etc. These compounds areeffective as both injectable and oral deliverable pharmaceuticalcompositions. Such compositions are prepared in a manner well known inthe pharmaceutical art and comprise at least one active compound.

The pharmaceutical compositions are formulated in the presence of apharmaceutically acceptable carrier. In making the compositions of thisinvention, the active ingredient is usually mixed with an excipient,diluted by an excipient or enclosed within such a carrier which can bein the form of a capsule, sachet, paper or other container. When theexcipient serves as a diluent, it can be a solid, semi-solid, or liquidmaterial, which acts as a vehicle, carrier or medium for the activeingredient. Thus, the compositions can be in the form of tablets, pills,powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions,solutions, syrups, etc., containing, for example, up to 10% by weight ofthe active compound, soft and hard gelatin capsules, suppositories,sterile injectable solutions, and sterile packaged powders.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, sterile water, syrup, and methylcellulose. The formulations can additionally include: lubricating agentssuch as talc, magnesium stearate, and mineral oil; wetting agents;emulsifying and suspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; sweetening agents; and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

The 1-galactose derivatives of this invention can also be administeredin the form of pro-drugs, i.e., as derivatives which are converted intoa biologically active compound of formula I in vivo. Such pro-drugs willtypically include compounds of formula I in which at least one of R^(a),R^(b), R^(c), or R^(d) is a biologically liable group, such as --C(O)R²or --P(O)(OR³)₂, where R² and R³ are as defined above.

The following synthetic and biological examples are offered toillustrate this invention and are not to be construed in any way aslimiting the scope of this invention. Unless otherwise stated, alltemperatures are in degrees Celsius.

EXAMPLES

In the examples below, the following abbreviations have the followingmeanings. If an abbreviation is not defined, it has its generallyaccepted meaning.

    ______________________________________    Å = angstroms    bd = broad doublet    bs = broad singlet    BSA = bovine serum albumin    d = doublet    dd = doublet of doublets    DMAP = dimethylaminopyridine    eq. = equivalents    g = grams    L = liter    m = multiplet    meq = milliequivalent    mg = milligram    mL = milliliter    mmol = millimole    N = normal    OPD = o-phenylenediamine    PBS = phosphate buffered saline at pH 7.2    q = quartet    quint. = quintet    s = singlet    t = triplet    TFA = trifluoroacetic acid    THF = tetrahydrofuran    TLC = thin layer chromatography    Tween 20 = polyoxyethylenesorbitan monolaurate    μL = microliter    ______________________________________

¹ H-Nmr spectra were recorded with a Brueker AM-360 spectrometer andMALDI-TOF mass spectra were recorded with a -HP G2020A (LD-TOF)instrument. Optical rotations were measured with a Perkin-Elmer 241polarimeter. Reactions were monitored by TLC on Silica Gel FG254 (E.Merck, Darmstadt, Germany).

Example A Solid-Phase Extraction of Lauroylated Intermediates

As indicated in the following examples, certain lauroylated reactionintermediates were purified by solid-phase extraction. In thispurification procedure, the reaction mixture is concentrated,re-dissolved in methanol, and applied onto C18 silica (Waters Prep C18,125 Å, 1 g per 20 mg lauroylated carbohydrate). The C18 silica is thenwashed with methanol (10 mL/g C18 silica) and the product is eluted withpentane (10 mL/g C18 silica). For L-arginine containing compounds, thereaction mixture is concentrated, re-dissolved in 70% methanol andapplied onto C18 silica. The C18 silica is then washed with 70% methanoland the product is eluted with methanol. The resulting product containsno residual reagents as determined by TLC, ¹ H-nmr, or MALDI-TOF massspectroscopy.

Example B Synthesis of 1,2,3,4,6-Penta-O-lauroyl-α-D-galactopyranose 1

To a suspension of galactose (3.78 g, 21.0 mmol), pyridine (50 mL), and4dimethylaminopyridine (cat.) in pentane (150 mL) under argonatmosphere, was added lauroyl chloride (50 mL, 210 mmol) at -78° C. Themixture was allowed to reach ambient temperature. The resulting whiteslurry slowly dissolved and a fine precipitate of pyridiniumhydrochloride formed. After 40 h, the pyridinium hydrochloride wasfiltered off and the pentane solution was concentrated. Columnchromatography (SiO₂, pentane/EtOAc 9:1) gave 1 (16.0 g, 70% yield),α!_(D) ²⁵ +39° (c 0.9, CHCl₃). ¹ H-Nmr data (CHCl₃): δ 6.39 (d, 1H, J2.4 Hz, H-1), 5.51 (br s, 1H, H-4), 5.35 (m, 2H, H-2 and H-3), 4.32 (brt, 1H, J 6.6 Hz, H-5), 4.08 (d, 2H, J 6.6 Hz, H-6a and H-6b), 2.39,2.38, 2.30, 2.26 (4 t, 2H each, J 7.5 Hz, --CH₂ CO--), 2.21 (m, 2H,--CH₂ CO--), 0.88 (t, 15H, J 7.0 Hz, --CH3). Anal. Calcd for C₆₆ H₁₂₂O₁₁ : C, 72.2; H, 11.3. Found: C, 72.6; H, 11.5.

Example C Synthesis of1-S-Acetyl-2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranose (2)

Method 1: To compound 1 (from Example B, 1 g, 0.91 mmol) and thiolaceticacid (0.4 mL, 9.1 mmol) in dry dichloromethane (5 mL) under argon at 0°C., was added boron trifluoride etherate (1.7 mL, 13.6 mmol). Thecold-bath was removed after 10 min and after 24 h the mixture wasdiluted with dichloromethane, washed with saturated sodium bicarbonate,dried over sodium sulfate, and concentrated. Column chromatography(SiO₂, pentane/Et2O/EtOAc 9:1:1) gave 2 (0.60 g, 70% yield).

Method 2: To compound 1 (from Example B, 276.5 mg, 0.253 mmol) in drytetrahydrofuran (2.0 mL) under argon, was added benzylamine (27.9 μL,0.255 mmol). The mixture was concentrated after 70 h. The residue wasdissolved in dry dichloromethane (4.0 mL) under argon and thentrichloroacetonitrile (250 μL, 2.5 mmol) and 1,8-diazabicyclo5.4.0!undec-7-ene (30 μL, 0.2 mmol) were added. The mixture wasconcentrated after 3 h and the residue was flashed through a shortcolumn (SiO₂, pentane/EtOAc 19:1), then concentrated. To the residue indry dichloromethane (3.5 mL) under argon, was added thiolacetic acid (1mL). After 96 h, the reaction mixture was concentrated and the residuewas purified by column chromatography (SiO₂, pentane, EtOAc 19:1) togive 2 (90 mg, 37% yield), α!_(D) ²⁵ 21° (c 1, CHCl₃). ¹ H-Nmr data(CHCl₃): δ 5.47 (d, 1H, J 3.4 Hz, H-4), 5.32 (t, 1H, J 10.0 Hz, H-2),5.25 (d, 1H, J 10.0 Hz, H-1), 5.12 (dd, 1H, J 3.4 and 10.0 Hz, H-3),4.08 (m, 3H, H-5, H-6a and H-6b), 2.14-2.43 (m, 8H, --CH₂ CO--), 2.37(s, 3H, -SAc), 0.88 (t, 15H, J 7.0 Hz, --CH3). Anal. Calcd for C₅₆ H₁₀₂O₁₀ S: C, 69.5; H, 10.6; S, 3.3. Found: C, 69.4; H, 10.8; S, 3.5.

Method 3: To compound 1 (20.0 g, 18.2 mmol) and thioacetic acid (5.0 mL,1.9 eq.) in dry dichloromethane (300 mL) under argon, was addedtrimethylsilyl trifluoromethanesulfonate (5.0 mL, 0.5 eq.) at 0° C. Thecold-bath was immediately removed and after 48 h the mixture was dilutedwith dichloromethane, washed with saturated sodium hydrogen carbonate,dried (Na₂ SO₄), and concentrated. Column chromatography (SiO₂,pentane/EtOAc 20:1) gave 2 (13.7 g, 77%), α!_(D) ²⁵ +21° (c 1, CHCl₃). ¹H-Nmr data (CHCl₃): d 5.47 (d, 1H, J 3.4 Hz, H4), 5.32 (t, 1H, J 10.0Hz, H-2), 5.25 (d, 1H, J 10.0 Hz, H-1), 5.12 (dd, 1H, J 3.4 and 10.0 Hz,H-3), 4.08 (m, 3H, H-5, H-6a and H-6b), 2.14-2.43 (m, 8H, --CH₂ CO--),2.37 (s, 3H, -SAc), 0.88 (t, 15H, J 7.0 Hz, --CH₃). Anal. Calcd for C₅₆H₁₀₂ O₁₀ S: C, 69.5; H, 10.6; S, 3.3. Found: C, 69.4; H, 10.8; S, 3.5.

Example C' Synthesis of1-S-Acetyl-2,3,4,6-tetra-O-lauroyl-1-thio-α-D-galactopyranose

Method 1: To compound 1 (20.0 g, 18.2 mmol) and thioacetic acid (27.0mL, 20 eq.) in dry toluene (80 mL) under argon was added tin (IV)chloride (21.3 mL) dropwise at room temperature. The reaction wasmonitored by Tlc carefully. After 1 h, 600 mL of 1M aqueous HCl wasadded to the vigorously stirred mixture and the resulting mixture wasfiltered through Celite to remove the emulsion of tin salts. The mixturewas diluted with pentane (800 mL), washed with water (2×400 mL),saturated sodium hydrogen carbonate (300 mL) and water (300 mL), driedwith Na₂ SO₄ and concentrated. The residue was purified by columnchromatography three times (SiO₂, pentane/EtOAc 20:1, 30:1, 40:1) togive 1-S-acetyl-2,3,4,6-tetra-O-lauroyl-1-thio-α-D-galactopyranose (3.65g, 21%). ¹ H-Nmr data (CHCl₃): δ 6.26 (d, 1H, J 5.5 Hz, H-1), 5.47 (dd,1H, J 11.0 Hz, 5.5 Hz, H-2), 5.46 (d, 1H, J 3.5 Hz, H-4), 5.04 (dd, 1H,J 11.0 Hz, 3.5 Hz, H-3), 4.17 (t, 1H, J 6.5 Hz, H-5), 4.06 (d, 2H, J 6.5Hz, H-6a and H-6b), 2.38 (t, 8H, J 7.0 Hz, --COCH₂ --), 2.40 (s, 3H,-SAc), 0.87 (t, 15H, J 7.0 Hz, --CH₃).

Method 2: To compound 1 (25.0 g, 22.9 mmol) and thioacetic acid (8.5 mL,114.5 mmol) in dry dichloromethane (100 mL) under argon, was addedtrimethylsilyl trifluoromethanesulfonate (5.6 mL, 45.8 mmol) at roomtemperature. After 20 h, the mixture was diluted with dichloromethane(600 mL), washed with saturated sodium hydrogen carbonate (250 mL) andwater (2×200 mL), dried with Na₂ SO₄ and concentrated. The residue waspurified by column chromatography three times (SiO₂, pentane/EtOAc 20:1,30:1, 40:1) to give1-S-acetyl-2,3,4,6tetra-O-lauroyl-1-thio-α-D-galactopyranose (1.59 g,7.2%).

Example D General Procedure for Michael Additions and α-HalocarbonylSubstitutions

To compound 2 (1 mmol) and an electrophile (1.2 mmol) in drydichloromethane (8 mL) under argon, was added Et₂ NH (4 mL). After 1-3h, the mixture was concentrated and the residue was purified by columnchromatography on SiO₂ by eluting with pentane/EtOAc. The products werecharacterized with ¹ H-nmr spectroscopy and MALDI-TOF mass spectroscopy.

Example E General Procedure for Reduction to Alcohols

To the product from Example D (100 μmol) in dry tetrahydrofuran (2.0 mL)and isopropanol (0.7 mL) under argon atmosphere, was added NaBH₄ (150μmol). After 0.5-3 h, the mixture was concentrated and the residue waspurified according to the solid-phase extraction procedure of Example A.The product alcohols were characterized with ¹ H-nmr spectroscopy andMALDI-TOF mass spectroscopy.

Example F General Procedure for Reductive Amination to a Primary Amine

Method 1: To the product from Example D (100 μmol) and ammonium acetate(75 mg, 1 mmol) in dry methanol (2.3 mL) and tetrahydrofuran (0.9 mL)under argon, was added NaCNBH₃ (100 μmol). After 1-72 h, the mixture wasconcentrated and the residue purified according to the solid-phaseextraction procedure of Example A. The product amines were characterizedwith ¹ H-nmr spectroscopy and MALDI-TOF mass spectroscopy.

Method 2: The product from Example D (200 mg, 0.198 mmol) and dry NH₄OAc (30 mg, 0.4 mmol) were stirred in dry MeOH (6 mL), dry1,2dichloroethane (6 mL), and trimethyl orthoformate (1 mL) under argonfor 24 h (until TLC analysis showed that most of the starting materialwas consumed). NaBH₄ (10 mg, 0.26 mmol) was added and after 1 h themixture was concentrated. The residue was purified according to thesolid-phase extraction procedure of Example A to provide the primaryamine (containing traces of the corresponding alcohol). This mixture wasdissolved in pentane/EtOAc (1:1) and applied onto a Waters Sep-Pak PlusLongbody SiO₂ cartridge. The cartridge was washed with pentane/EtOAc(1:1, 20 mL) (to remove the corresponding alcohol), followed by elutionwith toluene/EtOH (9:1, 30 mL) to afford the primary amine.

Example G General Procedure for the Preparation of Mesylates

To the alcohol from Example D (0.3 mmol) in dry tetrahydrofuran (2 mL)and dry pyridine (4 mL) under an argon atmosphere was addedmethanesulfonyl chloride (0.5 mL). After 12-24 h, the mixture was washedwith 0.5M HCl and extracted with pentane. The pentane extracts wereconcentrated and the residue was purified on C18-silica gelchromatography to afford the mesylate derivative.

Example H General Procedure for the Preparation of Azido Compounds

To the mesylate from Example G (0.2 mmol) in dry DMF (8 mL) and dry THF(3 mL) under an argon atmosphere at 60° C. was added sodium azide (5mmol) and 18-crown-6 (180 mg). After 2 hours, the reaction mixture wasconcentrated and the residue was purified on C18-silica. In some cases,the product was re-chromatographed with silica gel using pentane/EtOAc(9:1) as the eluant to afford the azido derivative.

Example I General Procedure for Reduction of Azido Groups to PrimaryAmines

To a solution of the azido compound from Example H (15 μmol) in dryisopropanol (1 mL) and dry ethanol (1 mL) under an argon atmosphere, wasadded NaBH₄ (15 μmol) and NiCl₂ (30 μmol). After 1 hour, the reactionmixture was neutralized with acetic acid (1 drop), concentrated andpurified on C18-silica to afford the primary amine.

Example J General Procedure for Reductive Alkylation of Primary Amines

To the primary amine from Example F or I (6.8 μmol) in dry methanol (1mL) and dry dichloromethane (1 mL) under an argon atmosphere was addedan aldehyde or ketone (3.4 mmol) and sodium triacetoxyborohydride (47μmol). After 24-48 hours, toluene (1 mL) was added and the mixture wasconcentrated and the residue purified on C18-silica gel.

Example K General Procedure for Reductive Amination

To the product from Example D (0.1 mmol) and an amine (0.45 mmol) in drydichloromethane (2 mL), methanol (2 mL) and triethylorthoformate (1 mL)under argon, was added NaCNBH₃ (1 mmol). After 24 h, the mixture wasconcentrated and dissolved in toluene (1 mL) and purified on C18-silicagel (5 g).

Example L General Procedure for Deblocking 2,3,4,5-tetra-O-lauroyl1-Galactose Derivatives

To the O-lauroylated 1-galactose derivative (100 μmol) in dry methanol(7.1 mL) and dichloromethane (1.4 mL) under argon, was added methanolicsodium methoxide (1M, 50μL). After 1-24 h, the mixture was neutralizedwith Amberlite IR-50S (H⁺) resin, filtered and concentrated. The residuewas dissolved in dichloromethane/methanol 2:1 and applied to a WatersSepPak Plus Longbody SiO₂ cartridge. The cartridge was washed withdichloromethane/methanol (2:1) and then the product was eluted withdichloromethane/methanol/water (5:5:1) (20 mL) and concentrated. Theresidue was dissolved in water and applied onto a column of C18 silica(Waters Prep C18, 125 Å, 5 g). The C18 silica was washed with water (50mL) and then the product was eluted with methanol (50 mL). The resultingsecondary amines were characterized with 1H-nmr spectroscopy andMALDI-TOF mass spectroscopy.

Example B6H Synthesis of2,2-Dimethyl-4-(cyclobut-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile andcyclobutylamine as the primary amine. Mass spectra data was as follows:M (calcd.): 361.50; M (found): 361.6 (M+H⁺). Selected nmr data was asfollows: ¹ H-nmr (CD₃ OD): δ 4.315 (H-1), 4.300, 4.292, 4.290.

Example B6I Synthesis of2,2-Dimethyl-4-(3,3-dimethylcyclobut-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile and3,3-dimethylcyclobut-1-ylamine as the primary amine. Mass spectra datawas as follows: M (calcd.): 389.55; M (found): 392.2 (M+H⁺). Selectednmr data was as follows: ¹ H-nmr (CD₃ OD): δ 4.324 (H-1), 4.311, 4.305,4.294.

Example B6J Synthesis of2,2-Dimethyl-4-(cyclopent-1-ylamino)cyclopent-11-y1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile andcyclopentylamine as the primary amine. Mass spectra data was as follows:M (calcd.): 375.52; M (found): 376.6 (M+H⁺). Selected nmr data was asfollows: ¹ H-nmr (CD₃ OD): δ 4.322 (H-1), 4.310, 4.304, 4.295.

Example B6K Synthesis of2,2-Dimethyl-4-(cyclohex-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile andcyclohex-1-ylamine as the primary amine. Mass spectra data was asfollows: M (calcd.): 389.55; M (found): 391.2 (M+H⁺). Selected nmr datawas as follows: ¹ H-nmr (CD₃ OD): δ 4.319 (H-1), 4.310, 4.307, 4.293.

Example B6L Synthesis of2,2-Dimethyl-4-(4-methylcyclohex-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile and4-methylcyclohex-1-ylamine as the primary amine. Mass spectra data wasas follows: M (calcd.): 403.47; M (found): 404.8 (M+H⁺). Selected nmrdata was as follows: ¹ H-nmr (CD₃ OD): δ 4.333 (H-1), 4.312, 4.300,4.295.

Example B6Q Synthesis of2,2-Dimethyl-4-(3-methylcyclopent-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile and3-methylcyclopent-1-ylamine as the primary amine. Mass spectra data wasas follows: M (calcd.): 389.55; M (found): 390.7 (M+H⁺). Selected nmrdata was as follows: ¹ H-nmr (CD₃ OD): δ 4.383 (H-1), 4.325, 4.300,4.292.

Example B6R Synthesis of2,2-Dimethyl-4-(3,3-dimethylcyclopent-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile and3,3-dimethylcyclopent-1-ylamine as the primary amine. Mass spectra datawas as follows: M (calcd.): 4.295; M (found): 404.3 (M+H⁺). Selected nmrdata was as follows: ¹ H-nmr CD₃ OD): δ 4.322 (H-1), 4.305, 4.300,4.295.

Example B6T Synthesis of2,2-Dimethyl-4-(3-methylcyclohex-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

The title compound was prepared according to procedures D, K and L aboveusing 4,4-dimethylcyclopent-2-en-1-one as the electrophile and3-methylcyclohex-1-ylamine as the primary amine. Mass spectra data wasas follows: M (calcd.): 403.57; M (found): 404.8 (M+H⁺). Selected nmrdata was as follows: ¹ H-nmr (CD₃ OD): δ 4.326 (H-1), 4.313, 4.303,4.294.

Example 1 Synthesis of the Individual Diastereomers of2,2-Dimethyl-4-(cyclobut-1-ylamino)cyclopent-1-yl1-Thio-β-D-galactopyranoside

This example illustrates the preparation of individual diastereomers ofa compound of formula I.

Step A--Synthesis of(1R,S)-2,2-Dimethylcyclopentan-4-on-1-yl2,3,4,6-Tetra-O-lauroyl-1thio-.beta.-D-galactopyranoside:To 1-S-acetyl-2,3,4,6-tetra-O-lauryl-1-thio-β-D-galactopyranose (5 g, 5mmol) (from Example B above) and 4,4-dimethyl-2-cyclopenten-1-one (500mg, 4.45 mmol) in dry CH₂ Cl₂ (10 mL) under argon, was added Et₂ NH (6mL). After 3 h, the mixture was concentrated and purified by columnchromatography (SiO₂, pentane/EtOAc, 9:1) to give the title compound asa mixture of diastereomers (3.54 g, 66%).

Step B--Separation of the Diastereomers of(1R,S)-2,2-Dimethylcyclopentan-4-on-1-yl2,3,4,6-Tetra-O-lauroyl-1-thio-β-D-galactopyranoside: The twodiastereomers from Step A (5 g, 4.8 mmol) were separated by columnchromatography (SiO₂, pentane/EtOAc, 9:1) to give(1S)-2,2-dimethylcyclopentan-4-on-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (428.8 mg, 8%) and(1R)-2,2-dimethylcyclopentan-4-on-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (373.8 mg, 6%)along with a mixture of unresolved compounds (2.74 g, 52%).

Step C--Synthesis of (1S, 4RS)- and (1R,4RS)-2,2-Dimethyl-4-hydroxycyclopent-1-yl2,3,4,6-Tetra-O-lauroyl-1-thio-β-D-galactopyranoside: To each of thepurified diastereomers from Step B (in separate reaction flasks) (320mg, 0.3 mmol) in dry tetrahydrofuran (3 mL), methanol (0.5 mL) andisopropanol (2 mL) under argon atmosphere, was added NaBH₄ (0.12 mmol).After 30 min, AcOH (1 drop) is added and the mixtures were concentratedand the residues dissolved MeOH (2 mL) and added to a column of C-18silica (5 g). The columns were washed with MeOH (50 mL) and productseluted pentane (50 mL) to give (1S,4RS)-2,2-dimethyl-4-hydroxy-cyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (281 mg, 88%) and(1R, 4RS)-2,2-dimethyl-4-hydroxy-cyclopent-1-y2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (297 mg, 93%).

Step D--Synthesis of (1S, 4RS)- and (1R,4RS)-2,2-Dimethyl-4-O-methanesulfonyloxycyclopent-1-yl2,3,4,6-Tetra-O-lauroyI-1-thio-β-D-galactopyranoside: To each of the(1S, 4RS) and (1R, 4RS) mixtures from Step C (in separate reactionflasks) (280 mg, 0.3 mmol) in dry tetrahydrofuran (2 mL) and drypyridine (4 mL) under argon atmosphere, was added methanesulfonylchloride (0.5 mL). After 12 h, the mixtures were washed with 0.5 M HCland extracted with pentane. After concentration, the residues werepurified on C18-silica (5 g) as described in Step C to afford (1S,4RS)-2,2-dimethyl-4-O-methanesulfonyloxycyclopent-1-yl 2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (281 mg, 88%) and (5)(1R, 4RS)-2,2-dimethyl-4-O-methanesulfonyloxycyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (297 mg, 93%) aswhite solids after pentane evaporation.

Step E--Synthesis of (1S, 4R)-, (1S, 4S)-, (1R, 4S)- and (1R,4R)-2,2-Dimethyl-4-azidocyclopent-1-yl2,3,4,6-Tetra-O-lauroyl-1-thio-β-D-galactopyranoside: To the (1S, 4RS)and (1R, 4RS) mixtures from Step D (in separate reaction flasks) (250mg, 0.2 mmol) in dry DMF (8 mL) and dry THF (3 mL) under argonatmosphere at 60° C. was added NaN₃ (340 mg, 5 mmol) and 18 crown-6 (180mg). After 2 h, the mixtures were concentrated and purified on C18silica(5 g) as described in Step C. Re-chromatography (SiO₂, pentane/EtOAc,9:1) permitted the separation of diastereomers to give pure (1S,4R)-2,2-dimethyl-4-azidocyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (163 mg, 65%); (1S,4S)-2,2-dimethyl-4-azidocyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (29 mg, 9%); (1R,4S)-2,2-dimethyl-4-azidocyclopent-1-yl2,3,4,6tetra-O-lauroyl-1-thio-β-D-galactopyranoside (68 mg, 28%); and(1R, 4R)-2,2-dimethyl-4-azidocyclopent-1-yl2,3,4,6-tetra-O-lauroyl-1-thio-β-D-galactopyranoside (21 mg, 9%).

Step F--Synthesis of (1S, 4R)-, (1S, 4S)-, (1R, 4S)- and (1R,4R)-2,2Dimethyl-4-amino-cyclopent-1-yl2,3,4,6-Tetra-O-lauroyl-1-thio-β-D-galactopyranoside: To each of thefour diastereomers of 2,2-dimethyl-4-azidocyclopent-1-yl1-thio-β-D-galactopyranoside from Step E (5 mg, 15 μmol) in dryisopropanol (1 mL) and dry ethanol (1 mL) under argon atmosphere, wasadded NaBH₄ (15 μmol) and NiCl₂ (30 μmol). After 1 h, the mixtures wereneutralized with AcOH (1 drop), concentrated and purified on C18-silica(2 g) as described in Step C to give (1S, 4R)-, (1S, 4S)-, (1R, 4S)- and(1R, 4R)-2,2-dimethyl-4-aminocyclopent-1-yl 1-thio-β-D-galactopyranoside(each 5 mg; quant.).

Step G--Synthesis of (1S, 4R)-, (1S, 4S)-, (1R, 4S)- and (1R,4R)-2,2-Dimethyl-4-(cyclobut-1-ylamino)cyclopent-1-yl2,3,4,6-Tetra-O-lauroyl-1-thio-β-D-galactopyranoside: To each of fourdiastereomers of 2,2-dimethyl-4-aminocyclopent-1-yl1-thio-β-D-galactopyranoside from Step F (in separate reaction flasks)(2 mg, 6.8 μmol) in dry methanol (1 mL) and dry dichloromethane (1 mL)under argon atmosphere, was added cyclobutanone (250 μL, 3.4 mmol) andsodium triacetoxyborohydride (10 mg, 47 μmol). After 24-48 h, toluene (1mL) was added and the mixture was concentrated and the residue purifiedon C18-silica as described in Step C to give 2.1-2.4 mg (quant.) eachof:

(1S, 4R)-2,2-dimethyl-4-(cyclobut-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside (B6HA); M (calcd.): 361.50; M (found):361.6 (M+H⁺); ¹ H-nmr (CD₃ OD): δ 4.292 (H-1);

(1S, 4S)-2,2-dimethyl-4-(cyclobut-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside (B6HB); M (calcd.): 361.50; M (found):361.6 (M+H⁺); ¹ H-nmr (CD₃ OD): δ 4.315 (H-1);

(1R, 4S)-2,2-dimethyl-4-(cyclobut-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside (B6HC); M (calcd.): 361.50; M (found):361.6 (M+H⁺); ¹ H-nmr (CD₃ OD): δ 4.300 (H-1);

(1R, 4R)-2,2-dimethyl-4-(cyclobut-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside (B6HD); M (calcd.): 361.50; M (found):361.6 (M+H⁺); ¹ H-nmr (CD₃ OD): δ 4.290 (H-1).

Example 2 Attachment of a Carboxyl-Containing 1-β-D-galactopyranoside toa Solid Support

To a 1-β-D-galactopyranoside of formula I having a carboxyl group on theA or B ring (2.1 mg, 4.5 μmol), silyl aminated Chromosorb P (449 mg,prepared as described in U.S. Pat. No. 4,137,401¹² and Westal et al.¹³),and hydroxybenzotriazole (1.3 mg, 9.4 μmol) in DMF (1 mL, dried over 4Amolecular sieves), is added diisopropylcarbodiimide (1.4 μL, 9.0 μmol).The beads are filtered off after 75 hours, washed with water, DMF, MeOH,and CH₂ Cl₂. To the resulting beads in MeOH (1.5 mL) is added aceticanhydride (0.5 mL) and after 16.5 hours, the beads are filtered andwashed with water, DMF, MeOH, CH₂ Cl₂, and pentane. Fine particles areremoved by suspending the beads in MeOH and decanting the supernatantrepeatedly. Drying under high-vacuum provides a product having the1-β-D-galactopyranoside covalently attached to the chromasorb P byformation of an amide linkage between amine group of the chromasorb Pand the carboxy group of the 1-thiogalactose derivative. Phenol/H₂ SO₄assay using the procedure described in M. Dubois et al.¹¹ can be used toshow the incorporation yield.

Example 3 Solid-Phase Synthesis of 1-Galactose Derivatives

The example illustrates the solid-phase synthesis of 1-galactosederivatives of formula I.

Step A--Synthesis of1-Dithioethyl-2,3,4,6-tetra-O-acetyl-galactopyranoside:1-Thio-2,3,4,6-tetra-O-acetyl-galactopyranoside (500 mg, 1.37 mmol) anddiethyl-N-ethyl-sulfenylhydrazodicarboxylate (360 mg, 2.0 mmol)(prepared as described in T. Mukaiyama¹⁴) are dissolved indichloromethane (14 mL) and stirred at room temperature. After 10 min,the solution is concentrated and column chromatography (SiO₂,hexane/ethylacetate 2:1) yields1-dithioethyl-2,3,4,6-tetra-O-acetyl-galactopyranoside (580 mg, quant)as a white solid (R_(f) 0.27 in hexanes/ethyl acetate (2:1)).

¹ H-NMR (360 MHz, CHCl₃): δ 1.30 (dd, 3H, J=7.4 Hz, CH₃), 1.96, 2.02,2.03, 2.13 (4 s, 12H, 4 CH₃ CO), 2.79 (ddd, 2H, J=7.4 Hz, J=7.4 Hz,J=1.3 Hz, CH₂), 3.94 (ddd, 1H, J₄,5 =1.0 Hz, J₅.6a =6.6 Hz, J₅,6b =7.6Hz, 5-H), 4.10 (ddd, 2H, 61-H, 6b-H), 4.51 (d, 1H, J₁,2 =10.0 Hz, 1-H),5.05 (dd, 1H, J₂,3 =10.0 Hz, J₃,4 =3.3 Hz, 3-H)), 5.38 (dd, 1H, J₁,2=10.0 Hz, J₃,3 =10.0 Hz, 2-H), 5.40 (dd, 1H, J₃,4 =3.3 Hz, J₄,5 =1.0 Hz,4-H); m/z calcd. for C₁₆ H₂₄ O₉ S₂ (M+Na) 447.1, found 447.0.

Step B--Synthesis of 1-Dithioethyl-β-D-galactopyranoside:1-Dithioethyl-2,3,4,6-tetra-O-acetyl-galactopyranoside from Step A (500mg, 1.18 mmol) is dissolved in dry methanol (10 mL) and treated withmethanolic sodium methoxide (1 M, 150 μL). After 2 h, the solution isneutralized with Amberlite 1R-120 (H⁺) resin, filtered and concentratedto give 1-dithioethyl-6-β-D-galactopyranoside as a white solid (300 mg,quant).

Step C--Coupling of 1-Dithioethyl-β-D-galactopyranoside to a Resin:1-Dithioethyl-6-β-D-galactopyranoside (200 mg, 780 μmol) is dissolved indry pyridine (8 mL). Trityl chloride-resin (1 g, 950 μmol tritylchloride resin, loading 0.95 mmol/g of active chlorine, polymer matrix:copolystyrene-1% DVB, 200-400 mesh, Novabiochem) and DMAP (5 mg) areadded and the mixture is heated for 24 h at 60° C. The resin is filteredoff, and washed successively with methanol, tetrahydrofuran,dichloromethane and diethyl ether (10 mL each) to afford1-dithioethyl-β-D-galactopyranoside covalently linked to the tritylresin through the hydroxyl group in the 6-position.

Step D--Generation of the Free Thiol on the Resin: The resin from Step C(50 mg) is swollen in dry tetrahydrofuran (1.5 mL). Dry methanol (300μL), dithiothreitol (74 mg) and triethylamine (180 μL) are added and themixture is shaken for 10 hours at room temperature. The resin isfiltered off and washed successively with methanol, tetrahydrofuran,dichloromethane and diethyl ether (10 mL/each). IR (of intact beads):2565 cm⁻¹ (SH stretch).

Step E--Michael Addition Reaction: The resin from Step D (50 mg) isswollen in dry N,N-dimethylformamide (1 mL) and thencyclohept-2-en-1-one (70 μl, 63 μmol) is added and the mixture is shakenat room temperature. After 2 hours, the resin is filtered off and washedsuccessively with methanol, tetrahydrofuran, dichloromethane and diethylether (10 mL each).

Step F--Reductive Amination: The resin from Step E (50 mg) is swollen indichloromethane (1 mL). An aminocycloalkane (75 mg, 447 μmol), sodiumsulfate (100 mg), sodium triacetoxyborohydride (63 mg, 297 μmol) andacetic acid (10 μL) are added at room temperature under argon atmosphereand the mixture shaken for 24 hours. The resin is then filtered off andwashed successively with water, methanol, tetrahydrofuran anddichloromethane.

Step G--Cleavage from the 1-Thiogalactose Derivative from the Resin: Theresin from Step F (50 mg) is shaken with trifluoroacetic acid (1 mL) andtriisopropylsilane (20 μL) in dichloromethane (2 mL) at roomtemperature. After 3 hours, the resin is removed by filtration andwashed with dichloromethane (10 mL). After adding toluene (10 mL), thesolution is concentrated, then co-evaporated twice with toluene. Theresidue is dissolved in water (1 mL) and applied onto two C₁₈-Sep-Pak-cartridges (Waters Sep-Pak Plus). The C₁₈ silica is washed withwater (4 mL) and the final product is eluted with 20% methanol andconcentrated.

Example 4 Synthesis ofNα-(3-(β-D-Galactopyranosyl)cyclohept-1-yl)-L-leucine

Step A--Synthesis of Cycloheptan-1-on-3-yl2,3,4,6-Tetra-O-benzoyl-β-D-galactopyranoside:O-(2,3,4,6-tetra-O-benzoyl-β-galactopyranosyl) trichloroacetimidate (578mg, 780 μmol) and 3-hydroxycycloheptan-1-one (100 mg, 780 μmol) (whichwas prepared in two steps from cyclohept-2-en-1-one by the procedures ofE. Hasegawa⁸ and H. Paulson⁹ were dissolved in dry diethyl ether (10mL). Trimethyl trifluoromethanesulfonate (TMSOTf) (300 μmL, 0.1M) isadded at room temperature. After 1 h, the reaction is quenched withtriethylamine (100 μmol) and the mixture is concentrated. Columnchromatography (SiO₂, toluene/ethyl acetate 16:1 to 10:1) gives thetitle compound (385 mg, 70%) as a white foam.

Step B--Synthesis ofNα-(3-(2,3,4,6-Tetra-O-benzoyl-β-D-galactopyranosyl)cyclohept-1-yl)-L-leucinetert-Butyl Ester: To the product from Step A (50 mg, 71 μmol) andL-leucine tert-butyl ester dihydrochloride (80 mg, 354 μmol) indichloromethane (700 μL) and methanol 700 μL) was added sodiumcyanoborohydride (2 mg). After 48 h, the mixture was concentrated andpurified by column chromatography (SiO₂, toluene/ethyl acetate, 8:1) togive the title compound (48 mg, 78%).

Step C--Synthesis ofNα-(3-(β-D-Galactopyranosyl)cyclohept-1-yl)-L-leucine: The product fromStep B (12 mg, 14 μmol) was dissolved in dry methanol (1 mL) and treatedwith methanolic sodium methoxide (20 μL, 1M). After 16 h at 45° C., thesolution is neutralized with Amerlite IR-120 (H⁺) resin, filtered andconcentrated. The residue was then dissolved in aqueous lithiumhydroxide (0.1M, 500 μL). After 2 h, the mixture was neutralized withAmberlite IRC-50s (H⁺) resin, filtered and concentrated. The residue wasdissolved in water (500 μL) and applied to a C₁₈ -Sep-Pak cartridge(Waters Sep-Pak-plus). The C₁₈ column is washed with water (500 μL),whereafter the final product is eluted with a gradient of 10% to 30%methanol/water anc concentrated. After freeze drying from 1 mL of water,the title compound was obtained as a white powder (3 mg, 83%). ¹ H-Nmr(360 MHz, CD₃ OD) (selected signals): δ 0.84-1.08 (m, 6H, 2CH₃),5.31-5.37 (m, 1H, 1-H). This compound could be further cyclized toprovide a compound of this invention using conventional reagents andconditions.

By using a variety of cyclic amines in the above reactions, numerousother 1-galactose derivatives of this invention could be prepared.

Example 5 Inhibition of Heat-Labile Enterotoxin Binding to G_(D1b)

Using this example, the 1-galactose derivatives of formula I above couldbe tested for their ability to inhibit the binding of heat-labileenterotoxin from E. coli to ganglioside G_(D1b). The bioassay isconducted using the procedure described by A. M. Svennerholm¹⁵ exceptthat ganglioside G_(D1b) is used instead of ganglioside G_(M1). Thecompounds of Examples B6H, B6I, B6J, B6K, B6L, B6Q, B6R, and B6T areexpected to inhibit binding of heat-labile enterotoxin to gangliosideG_(D1b) by at least 20% in this assay.

Example 6 Inhibition of Cholera Toxin Binding to G_(D1b)

In this example, 1-galactose derivatives of formula I above were testedfor their ability to inhibit the binding of cholera toxin to gangliosideG_(D1b). This bioassay was conducted using the following modification ofthe procedure described by A. M. Svennerholm¹⁵.

On day 1, microtiter plates (C96 Maxisorp) were coated with 100 μL of 1mg/mL G_(D1b) (disialoganglioside G_(D1b), MW=2127, Fluka) in PBS perwell and incubated overnight at 37° C.

On day 2, the samples to be tested were diluted in BSA-Tween-PBS (0.1%BSA and 0.05% Tween-20 in PBS; Sigma). A total of 500 μL of eachsolution was prepared so that each point could be measured inquadruplicate. A concentration curve of 10, 20 and 30 ng/mL of CTB5-HRP(CT-B5 conjugated to HRP, Sigma, lyophilized, diluted in Tween-PBS) wasprepared. For the inhibition experiments, 20 ng/mL of CTB5-HRP was used.The samples were then incubated for 2 hours at room temperature. Afterincubation, the plates were emptied and unattached ganglioside wasremoved by washing the plates 2 times with 200 μL PBS per well.Additional binding sites on the plastic surface were then blocked byincubating the plates with 200 μL of 1% BSA in PBS per well for 30minutes at 37° C. The plates were then emptied and unattached BSA wasremoved by washing the plates 3 times with 200 μL of 0.05% Tween 20-PBSper well. Samples (100 μL) were added to 4 different wells and incubatedfor 30 minutes at room temperature. The plates were emptied andunattached BSA was removed by washing the plates 3 times with 200 μL of0.05% Tween 20-PBS per well.

A substrate solution was freshly prepared for each ELISA. Each solutioncontained 10 mg of o-phenylenediamine (Sigma), 5 mL of 0.1M sodiumcitrate (filter sterile or autoclaved), 5 mL of 0.1M citric acid (filtersterile or autoclaved) and 4 mL of 30% H₂ O₂. (Gloves should be wornsince o-phenylenediamine is carcinogenic). The substrate solution (100μL) was then added to each well and incubated for 30 minutes at roomtemperature. After incubation, the OD₄₅₀ was recorded. Under theconditions of the assay, D-galactose had an IC₅₀ of 30 mM.

The compounds of Examples B6H, B6I, B6J, B6K, B6L, B6Q, B6R, and B6Twere tested in this bioassay. All of the compounds tested inhibitedbinding of cholera toxin to ganglioside G_(D1b) by at least 10%.

Example 7 Neutralization of the Cytotonic Activity of CT and LT

This example illustrates how the solid support material of Example 2could be tested for its ability to neutralize the cytotonic activity ofCT and LT. The cytotonic activity of CT and LT is measured by the use ofChinese hamster ovary cells (CHO) that are maintained in Hams F12 mediasupplemented with 10% fetal bovine serum (FBS) in an atmosphere of 5%CO₂ at 37° C. Toxin samples are diluted 1:5 in Hams media and filtersterilized through 0.22 micron syringe filters. Samples are then serial5-fold diluted in media and 100 μL of each dilution is added to wellswith confluent monolayers of CHO cells and incubated for 24 h at 37° C.(under 5% CO₂). Each sample is analyzed two times. Cytotonic effects arereadily visible after 24 h incubation by comparing wells with controlsthat do not contain toxin. After 24 h, the cells are fixed with 95%methanol and stained with Geimsa stain. Toxin containing samples fromneutralization experiments are treated in an analogous fashion exceptthat the percent neutralization is determined by comparing the endpointdilutions of samples with and without the solid support material ofExample 2.

A solution containing purified CT or LT (2, 10 or 20 μg in 1 mL PBS) isadded to the solid support material of Example 2 (20 mg) in 1.5 mLmicrocentrifuge tubes and incubated at room temperature for 1 h on anend-over rotator. After incubation, the solid support material isallowed to settle to the bottom of the tubes and the supernatants arecarefully removed. The supernatants are added to CHO cells and thecytotonic endpoint determined after incubation for 24 h as describedabove. The extent of reduction in the endpoint in the presence of thesolid support material is determined by comparing with controls in whichsolid support material is not added.

A solid support material of Example 2 is expected to neutralized morethan 90% of CT and LT activity, i.e., less than 10% toxin activity willremain.

Example 8 Inhibition of Colonization Factor Antigens (CFA pili) Bindingto Glycophorin

This example illustrates how the 1-galactose derivatives of formula Iabove could be tested for their ability to inhibit CFA pili binding toglycophorin. Bacterial surface adhesion antigens such as CFA pili are avirulence factor expressed by certain enteric pathogens, includingenterotoxigenic E. coli. These pili are important factors in bacterialattachment to cell surface receptors. Accordingly, inhibition of CFApili binding is a useful test to determine whether a compound willinhibit the binding of a pathogenic microorganism to cell surfacereceptors.

Binding assays are done by coating microtitre wells with 50 μL ofglycophorin (10 μg/mL) in PBS for 2 h at 37° C. The solution is removedby aspiration and replaced with 100 μL of 1% BSA in PBS containing 0.05%Tween 20 (PBST) and incubated at 37° C. for an additional 1 h. Themicrotitre wells are washed three times with 200 μL of PBST and thenreplaced with biotinylated CFA I (5 μg/mL) in 50 μL of PBS containing0.05% BSA. After incubating for 2 h at 37° C., the binding reaction isstopped by aspirating the solutions and the plate is washed with PBST(3×200 μL). Avidin-peroxidase (50 μL of a 1/3000 dilution of a 1 mg/mLsolution in PBST containing 0.05% BSA) is added and the plates areincubated for an additional 1 h. After washing the wells as describedabove, 100 μL of the substrate solution (0.42 mM tetramethylbenzidine(TMB) in 0.1 M sodium citrate buffer, pH 6.0, containing 0.5 μM ureaperoxide) is added and the plates are incubated for 10 min at ambienttemperature and the enzyme reaction stopped by adding 50 μL of 2N H₂SO₄. Binding assays are done in triplicate and background binding ismeasured in wells coated with BSA only.

Binding inhibition assays are done using oligosaccharide analogs at aconcentration of 1 mg/mL in PBS. Inhibitors are pre-incubated withbiotinylated CFA I pili (5 μ/mL) for 1 h at 37° C. prior to adding toglycophorin-coated microtitre wells as outlined above.o-Nitrophenyl-β-D-galactose is utilized as a control inhibitor for theseexperiments.

The 1-galactose derivatives of Examples B6H, B6I, B6J, B6K, B6L, B6Q,B6R, and B6T are expected to inhibited CFA I pili binding to glycophorinin this assay.

From the foregoing description, various modifications and changes in thecomposition and method will occur to those skilled in the art. All suchmodifications coming within the scope of the appended claims areintended to be included therein.

I claim:
 1. A compound of formula I: ##STR4## wherein A is selected fromthe group consisting of arylene, cycloalkylene, cycloalkenylene,heteroarylene and divalent heterocyclic;B is selected from the groupconsisting of cycloalkyl, cycloalkenyl and heterocyclic; Y is selectedfrom the group consisting of oxygen, sulfur, --S(O)-- and --SO₂ --; Y'is selected from the group consisting of oxygen, sulfur, --S(O)--, --SO₂--, alkylene, substituted alkylene, and --NR¹ --, wherein R¹ is selectedfrom the group consisting of hydrogen, alkyl and acyl; and R^(a), R^(b),R^(c) and R^(d) are each independently selected from the groupconsisting of hydrogen; sulfate; --C(O)R², wherein R² is selected fromthe group consisting of alkyl, alkenyl, alkaryl, alkoxyalkyl, aryl,cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic and thioalkoxyalkyl;and --P(O)(OR³)₂, wherein each R³ is independently selected from thegroup consisting of hydrogen, alkyl, alkenyl, alkaryl, alkoxyalkyl,aryl, cycloalkyl, cycloalkenyl, heteroaryl, heterocyclic andthioalkoxyalkyl; on pharmaceutically acceptable salts thereof.
 2. Acompound of claim 1 wherein the compound of formula I is an α-anomer. 3.A compound of claim 1 wherein the compound of formula I is a β-anomer.4. A compound of claim 1 wherein A is a cycloalkylene group having 5 to8 carbon atoms.
 5. A compound of claim 4 wherein A is selected from thegroup consisting of cyclopentylene, methylcyclopentylene,dimethylcyclopentylene, cyclohexylene, methylhexylene,dimethylcyclohexylene and cycloheptylene.
 6. A compound of claim 1wherein B is a cycloalkyl group having 4 to 8 carbon atoms.
 7. Acompound of claim 6 wherein B is selected from the group consisting ofcyclobutyl, methylcyclobutyl, dimethylcyclobutyl, cyclopentyl,methylcyclopentyl, dimethylcyclopentyl, cyclohexyl, dimethylcyclohexyland cycloheptyl.
 8. A compound of claim 7 wherein B is cyclobutyl ordimethylcyclobutyl.
 9. A compound of claim 1 wherein Y is sulfur.
 10. Acompound of claim 1 wherein Y is oxygen.
 11. A compound of claim 1wherein Y' is --NH--.
 12. A compound of claim 1 wherein R^(a), R^(b),R^(c) and R^(d) are each hydrogen.
 13. A compound selected from thegroup consisting of:2,2-dimethyl-4-(cyclobutylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside 2.2-dimethyl-4-(3,3-dimethylcyclobut-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(cyclopentylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(3-methylcyclopent-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(3,3-dimethylcyclopent-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(cyclohexylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(3-methylcyclohex-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(4-methylcyclohex-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside and pharmaceutically acceptable saltsthereof.
 14. A pharmaceutical composition comprising from 1 to 99 weightpercent of a pharmaceutically acceptable carrier and from 1 to 99 weightpercent of at least one compound of formula I: ##STR5## wherein A isselected from the group consisting of arylene, cycloalkylene,cycloalkenylene, heteroarylene and divalent heterocyclic;B is selectedfrom the group consisting of cycloalkyl, cycloalkenyl and heterocyclic;Y is selected from the group consisting of oxygen, sulfur, --S(O)-- and--SO₂ --; Y' is selected from the group consisting of oxygen, sulfur,--S(O)--, --SO₂ --, alkylene, substituted alkylene, and --NR¹ --,wherein R¹ is selected from the group consisting of hydrogen, alkyl andacyl; and R^(a), R^(b), R^(c) and R^(d) are each independently selectedfrom the group consisting of hydrogen; sulfate; --C(O)R², wherein R² isselected from the group consisting of alkyl, alkenyl, alkaryl,alkoxyalkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl, heterocyclicand thioalkoxyalkyl; and --P(O)(OR³)₂, wherein each R³ is independentlyselected from the group consisting of hydrogen, alkyl, alkenyl, alkaryl,alkoxyalkyl, aryl, cycloalkyl, cycloalkenyl, heteroaryl, heterocyclicand thioalkoxyalkyl; on pharmaceutically acceptable salts thereof. 15.The pharmaceutical composition of claim 14 wherein the compound offormula I is an α-anomer.
 16. The pharmaceutical composition of claim 14wherein the compound of formula I is a β-anomer.
 17. The pharmaceuticalcomposition of claim 14 wherein A is a cycloalkylene group having 5 to 8carbon atoms.
 18. The pharmaceutical composition of claim 17 wherein Ais selected from the group consisting of cyclopentylene,methylcyclopentylene, dimethylcyclopentylene, cyclohexylene,methylhexylene, dimethylcyclohexylene and cycloheptylene.
 19. Thepharmaceutical composition of claim 14 wherein B is a cycloalkyl grouphaving 4 to 8 carbon atoms.
 20. The pharmaceutical composition of claim19 wherein B is selected from the group consisting of cyclobutyl,methylcyclobutyl, dimethylcyclobutyl, cyclopentyl, methylcyclopentyl,dimethylcyclopentyl, cyclohexyl, dimethylcyclohexyl and cycloheptyl. 21.The pharmaceutical composition of claim 20 wherein B is cyclobutyl ordimethylcyclobutyl.
 22. The pharmaceutical composition of claim 14wherein Y is sulfur.
 23. The pharmaceutical composition of claim 14wherein Y is oxygen.
 24. The pharmaceutical composition of claim 14wherein Y' is --NH--.
 25. The pharmaceutical composition of claim 14wherein R^(a), R^(b), R^(c) and R^(d) are each hydrogen.
 26. Apharmaceutical composition comprising from 1 to 99 weight percent of apharmaceutically acceptable carrier and from 1 to 99 weight percent ofat least one compound selected from the group consistingof:2,2-dimethyl-4-(cyclobutylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(3,3-dimethylcyclobut-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(cyclopentylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(3-methylcyclopent-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(3,3-dimethylcyclopent-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside 2.2-dimethyl-4-(cyclohexylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(3-methylcyclohex-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside2,2-dimethyl-4-(4-methylcyclohex-1-ylamino)cyclopent-1-yl1-thio-β-D-galactopyranoside and pharmaceutically acceptable saltsthereof.
 27. A method of ameliorating conditions associated with bindingof a toxin to its receptor in an animal which method comprisesadministering to said animal an effective amount of a pharmaceuticalcomposition of claim 14, wherein the compound of formula I inhibits thebinding of the toxin to its receptor.
 28. The method of claim 27 whereinthe toxin is heat-labile enterotoxin or cholera toxin.
 29. A method ofameliorating conditions associated with binding of a toxin to its cellsurface receptor in an animal which method comprises administering tosaid animal an effective amount of a pharmaceutical composition of claim26, wherein the compound inhibits the binding of the toxin to itsreceptor.
 30. The method of claim 29 wherein the toxin is heat-labileenterotoxin or cholera toxin.
 31. A method of ameliorating conditionsassociated with binding of an organism to its cell surface receptor inan animal which method comprises administering to said animal aneffective amount of a pharmaceutical composition of claim 14, whereinthe compound of formula I inhibits the binding of the organism to itscell surface receptor.
 32. The method of claim 31 wherein the organismis Vibrio cholerae or an enterotoxigenic strain of Escherichia coli. 33.A method of ameliorating conditions associated with binding of anorganism to its cell surface receptor in an animal which methodcomprises administering to said animal an effective amount of apharmaceutical composition of claim 26, wherein the compound of formulaI inhibits the binding of the organism to its cell surface receptor. 34.The method of claim 33 wherein the organism is Vibrio cholerae or anenterotoxigenic strain of Escherichia coli.
 35. A 1-galactosederivative-containing support comprising a support having covalentlybound thereto a plurality of at least one compound of formula I':##STR6## wherein A is selected from the group consisting of arylene,cycloalkylene, cycloalkenylene, heteroarylene and divalentheterocyclic;B is selected from the group consisting of cycloalkyl,cycloalkenyl and heterocyclic; Y is selected from the group consistingof oxygen, sulfur, --S(O)-- and --SO₂ --; Y' is selected from the groupconsisting of oxygen, sulfur, --S(O)--, --SO₂ --, alkylene, substitutedalkylene, and --NR³ --, wherein R³ is selected from the group consistingof hydrogen, alkyl and acyl; and R^(a), R^(b), R^(c) and R^(d) are eachindependently selected from the group consisting of hydrogen; sulfate;--C(O)R¹, wherein R¹ is selected from the group consisting of alkyl,alkenyl, alkaryl, alkoxyalkyl, aryl, cycloalkyl, cycloalkenyl,heteroaryl, heterocyclic and thioalkoxyalkyl; and --P(O)(OR²)₂, whereineach R² is independently selected from the group consisting of hydrogen,alkyl, alkenyl, alkaryl, alkoxyalkyl, aryl, cycloalkyl, cycloalkenyl,heteroaryl, heterocyclic and thioalkoxyalkyl; on pharmaceuticallyacceptable salts thereof;wherein one of A, B, R^(a), R^(b), R^(c) orR^(d) is covalently bound via a linking arm to the support.
 36. The1-galactose derivative-containing support of claim 35 wherein thesupport is a solid support.
 37. The 1-galactose derivative-containingsupport of claim 35 wherein the compound of formula I' is an α-anomer.38. The 1-galactose derivative-containing support of claim 35 whereinthe compound of formula I' is a β-anomer.
 39. The 1-galactosederivative-containing support of claim 35 wherein A is a cycloalkylenegroup having 5 to 8 carbon atoms.
 40. The 1-galactosederivative-containing support of claim 39 wherein A is selected from thegroup consisting of cyclopentylene, methylcyclopentylene,dimethylcyclopentylene, cyclohexylene, methylhexylene,dimethylcyclohexylene and cycloheptylene.
 41. The 1-galactosederivative-containing support of claim 35 wherein B is a cycloalkylgroup having 4 to 8 carbon atoms.
 42. The 1-galactosederivative-containing support of claim 41 wherein B is selected from thegroup consisting of cyclobutyl, methylcyclobutyl, dimethylcyclobutyl,cyclopentyl, methylcyclopentyl, dimethylcyclopentyl, cyclohexyl,dimethylcyclohexyl and cycloheptyl.
 43. The 1-galactosederivative-containing support of claim 42 wherein B is cyclobutyl ordimethylcyclobutyl.
 44. The 1-galactose derivative-containing support ofclaim 35 wherein Y is sulfur.
 45. The 1-galactose derivative-containingsupport of claim 35 wherein Y is oxygen.
 46. The 1-galactosederivative-containing support of claim 35 wherein Y' is --NH--.
 47. Apharmaceutical composition comprising from 1 to 99 weight percent of apharmaceutically acceptable carrier and from 1 to 99 weight percent of a1-galactose derivative-containing support comprising a support havingcovalently bound thereto a plurality of at least one compound of formulaI': ##STR7## wherein A is selected from the group consisting of arylene,cycloalkylene, cycloalkenylene, heteroarylene and divalentheterocyclic;B is selected from the group consisting of cycloalkyl,cycloalkenyl and heterocyclic; Y is selected from the group consistingof oxygen, sulfur, --S(O)-- and --SO₂ --; Y' is selected from the groupconsisting of oxygen, sulfur, --S(O)--, --SO² --, alkylene, substitutedalkylene, and --NR³ --, wherein R³ is selected from the group consistingof hydrogen, alkyl and acyl; and R^(a), R^(b), R^(c) and R^(d) are eachindependently selected from the group consisting of hydrogen; sulfate;--C(O)R¹, wherein R¹ is selected from the group consisting of alkyl,alkenyl, alkaryl, alkoxyalkyl, aryl, cycloalkyl, cycloalkenyl,heteroaryl, heterocyclic and thioalkoxyalkyl; and --P(O)(OR²)₂, whereineach R² is independently selected from the group consisting of hydrogen,alkyl, alkenyl, alkaryl, alkoxyalkyl, aryl, cycloalkyl, cycloalkenyl,heteroaryl, heterocyclic and thioalkoxyalkyl; on pharmaceuticallyacceptable salts thereof;wherein one of A, B, R^(a), R^(b), R^(c) orR^(d) is covalently bound via a linking arm to the support.
 48. Thepharmaceutical composition of claim 47 wherein the support is a solidsupport.
 49. The pharmaceutical composition of claim 47 wherein thecompound of formula I' is an α-anomer.
 50. The pharmaceuticalcomposition of claim 47 wherein the compound of formula I' is aβ-anomer.
 51. The pharmaceutical composition of claim 47 wherein A is acycloalkylene group having 5 to 8 carbon atoms.
 52. The pharmaceuticalcomposition of claim 51 wherein A is selected from the group consistingof cyclopentylene, methylcyclopentylene, dimethylcyclopentylene,cyclohexylene, methylhexylene, dimethylcyclohexylene and cycloheptylene.53. The pharmaceutical composition of claim 47 wherein B is a cycloalkylgroup having 4 to 8 carbon atoms.
 54. The pharmaceutical composition ofclaim 53 wherein B is selected from the group consisting of cyclobutyl,methylcyclobutyl, dimethylcyclobutyl, cyclopentyl, methylcyclopentyl,dimethylcyclopentyl, cyclohexyl, dimethylcyclohexyl and cycloheptyl. 55.The pharmaceutical composition of claim 54 wherein B is cyclobutyl ordimethylcyclobutyl.
 56. The pharmaceutical composition of claim 47wherein Y is sulfur.
 57. The pharmaceutical composition of claim 47wherein Y is oxygen.
 58. The pharmaceutical composition of claim 47wherein Y' is --NH--.
 59. A method of ameliorating conditions associatedwith binding of a toxin to its receptor in an animal which methodcomprises administering to said animal an effective amount of apharmaceutical composition of claim 47, wherein the compound of formulaI' inhibits the binding of the toxin to its receptor.
 60. The method ofclaim 59 wherein the toxin is heat-labile enterotoxin or cholera toxin.61. A method of ameliorating conditions associated with binding of anorganism to its cell surface receptor in an animal which methodcomprises administering to said animal an effective amount of apharmaceutical composition of claim 47, wherein the compound of formulaI' inhibits the binding of the organism to its cell surface receptor.62. The method of claim 61 wherein the organism is Vibrio cholerae or anenterotoxigenic strain of Escherichia coli.